Piperazine- and piperidine-derivatives as melanocortin receptor agonists

ABSTRACT

The present invention relates to melanocortin receptor agonists of formula I, which is useful in the treatment of obesity, diabetes and male and/or female sexual dysfunction

REFERENCE TO RELATED APPLICATION

This application is submitted as a United States national phase entry,pursuant to 35 U.S.C. §371, of PCT/US02/00515, filed on 23 Jan. 2002,which claims the benefit of U.S. provisional patent application Ser. No.60/263,471, filed 23 Jan. 2001.

FIELD OF THE INVENTION

The present invention relates to melanocortin receptor agonists, andmore particularly piperazine and piperidine derivatives as melanocortinreceptor agonists, which are useful for the treatment or prevention ofdiseases and disorders responsive to the activation of melanocortinreceptors.

BACKGROUND OF THE INVENTION

Pro-opiomelanocortin (POMC) derived peptides are known to affect foodintake. Several lines of evidence support the notion that the G-proteincoupled receptors (GPCRs) of the melanocortin receptor (MC-R) family,several of which are expressed in the brain, are targets of POMC derivedpeptides involved in the control of food intake and metabolism.

Evidence for the involvement of MC-R in obesity includes: i) the agouti(A^(vy)) mouse which ectopically expresses an antagonist of the MC-1R,MC-3R and MC-4R is obese, indicating that blocking the action of thesethree MC-Rs can lead to hyperphagia and metabolic disorders; ii) MC-4Rknockout mice (Huszar et al., Cell, 88:131–141, 1997) recapitulate thephenotype of the agouti mouse and these mice are obese; iii) the cyclicheptapeptide MC-1R, MC-3R, MC4R, and MC-5R agonist melanotanin-II(MT-II) injected intracerebroventricularly (ICV) in rodents, reducesfood intake in several animal feeding models (NPY, ob/ob, agouti,fasted) while ICV injected SHU-9119 (MC-3R, MC-4R antagonist; MC-1R andMC-5R agonist) reverses this effect and can induce hyperphagia; and iv)chronic intraperitoneal treatment of Zucker fatty rats with an α-NDP-MSHderivative (HP228) has been reported to activate MC-1R, MC-3R, MC-4R andMC-5R and to attenuate food intake and body weight gain over a 12 weekperiod.

Five MC-Rs have thus far been identified, and these are expressed indifferent tissues. MC-1R was initially characterized by dominant gain offunction mutations at the extension locus, affecting coat color bycontrolling phaeomelanin to eumelanin conversion through control oftyrosinase. MC-1R is mainly expressed in melanocytes. MC-2R is expressedin the adrenal gland and represents the ACTH receptor. MC-3R isexpressed in the brain, gut and placenta and may be involved in thecontrol of food intake and thermogenesis. MC-4R is uniquely expressed inthe brain and its inactivation was shown to cause obesity. (A. Kask, etal., “Selective antagonist for the melanocortin-4-receptor (HS014)increases food intake in free-feeding rats, Biochem. Biophys. Res.Commun., 245:90–93, 1998). MC-5R is expressed in many tissues includingwhite fat, placenta and exocrine glands. A low level of expression isalso observed in the brain. MC-5R knock out mice reveal reducedsebaceous gland lipid production (Chen et al., Cell, 91:789–798, 1997).

MC-4R appears to play role in other physiological functions as well,namely controlling grooming behavior, erection and blood pressure.Erectile dysfunction denotes the medical condition of inability toachieve penile erection sufficient for successful intercourse. The term“impotence” is often times employed to describe this prevalentcondition. Synthetic melanocortin receptor agonists have been found toinitiate erections in men with psychogenic erectile dysfunction (H.Wessells et al., “Synthetic Melanotropic Petide Initiates Erections inMen With Psychogenic Erectile Dysfunction: Double-Blind, PlaceboControlled Crossover Study,” J. Urol., 160: 389–393, 1998). Activationof melanocortin receptors of the brain appears to cause normalstimulation of sexual arousal. Evidence for the involvement of MC-R inmale and/or female sexual dysfunction is detailed in WO 00/74670.

Diabetes is a disease in which a mammal's ability to regulate glucoselevels in the blood is impaired because the mammal has a reduced abilityto convert glucose to glycogen for storage in muscle and liver cells. InType I diabetes, this reduced ability to store glucose is caused byreduced insulin production. “Type II Diabetes” or “non-insulin dependentdiabetes mellitus” (NIDDM) is the form of diabetes, which is due to aprofound resistance to insulin stimulating or regulatory effect onglucose and lipid metabolism in the main insulin-sensitive tissues,muscle, liver and adipose tissue. This resistance to insulinresponsiveness results in insufficient insulin activation of glucoseuptake, oxidation and storage in muscle and inadequate insulinrepression of lipolysis in adipose tissue and of glucose production andsecretion in liver. When these cells become desensitized to insulin, thebody tries to compensate by producing abnormally high levels of insulinand hyperinsulemia results. Hyperinsulemia is associated withhypertension and elevated body weight. Since insulin is involved inpromoting the cellular uptake of glucose, amino acids and triglyceridesfrom the blood by insulin sensitive cells, insulin insensitivity canresult in elevated levels of triglycerides and LDL which are riskfactors in cardiovascular diseases. The constellation of symptoms whichincludes hyperinsulemia combined with hypertension, elevated bodyweight, elevated triglycerides and elevated LDL is known as Syndrome X.

Spiropiperidine and piperidine derivates have been disclosed in U.S.Pat. No. 6,294,534 B1, WO 01/70337, WO 00/74679 and WO 01/70708 asagonists of melanocortin receptor(s), which can be used for thetreatment of diseases and disorders, such as obesity, diabetes andsexual dysfunction.

In view of the unresolved deficiencies in treatment of various diseasesand disorders as discussed above, it is an object of the presentinvention to provide novel piperazine derivatives, which are useful asmelanocortin receptor agonists to treat obesity, diabetes, and male andfemale sexual dysfunction.

SUMMARY OF THE INVENTION

The present invention relates to a compound of novel piperazine orpiperidine derivatives as melanocortin receptor agonists as shownformula I:

or a pharmaceutically acceptable salts or stereoisomers thereof, wherein

-   G is CR¹ or N;-   A is C₁–C₈ alkyl or C₃–C₇ cycloalkyl;-   L and L¹ are independently: hydrogen or together oxo;-   T is:

-   R is: when y is 1;    -   N(R⁸)₂,    -   NR⁸COR⁸,    -   NR⁸CON(R⁸)₂,    -   NR⁸C(O)OR⁸,    -   NR⁸C(R⁸)═N(R⁸),    -   NR⁸SO₂R⁸ or    -   NR⁸SO₂N(R⁸)₂;-   R is: when y is 0 or 1;    -   heterocyclyl, provided that when y is 0, a heteroatom is not        directly connected to oxygen or adjacent to a carbon that        connected to oxygen; and    -   wherein the heterocyclyl contains at least one nitrogen in the        ring and is optionally substituted with one to five substituents        independently selected from R⁸;-   R¹ is independently:    -   hydrogen, CONH(C₁–C₈ alkyl), C₁–C₈ alkyl, (D)phenyl, (D)C₃–C₇        cycloalkyl or oxo, provided that oxo is not attached to the same        carbon that attached to nitrogen which forms an amide bond when        G is N;-   R² is independently:    -   hydrogen,    -   halo    -   C₁–C₈ alkyl,    -   C₁–C₈ alkylsulfonyl,    -   (D)C₃–C₇ cycloalkyl or    -   C₁–C₄ haloalkyl;-   R³ is independently: aryl or thienyl;    -   wherein aryl and thienyl are optionally substituted with one to        three substituents selected from the group consisting of:    -   cyano, halo, C₁–C₈ alkyl, (D)C₃–C₇ cycloalkyl, C₁–C₄ alkoxy,        C₁–C₄ haloalkyl and C₁–C₄ haloalkyloxy;-   R⁴ is independently:    -   hydrogen, C₁–C₈ alkyl, C(O)R⁸, C(O)OR⁸, C₃–C₇ cycloalkyl or        (CH₂)_(n)O(C₁–C₈ alkyl), wherein n is 2–8;-   each R⁸ is independently:    -   hydrogen,    -   phenyl    -   C₁–C₈ alkyl,    -   C₁–C₈ alkylsulfonyl,    -   C(O)C₁–C₈ alkyl,    -   C(O)aryl, wherein aryl being phenyl or naphthyl,    -   SO₂-aryl, wherein aryl being phenyl or naphthyl,    -   (D)C₃–C₇ cycloalkyl or    -   (CH₂)_(n)C₁–C₄ haloalkyl, wherein n is 1–8;-   each R⁹ is independently:    -   hydrogen,    -   hydroxy,    -   (D)cyano,    -   halo,    -   C₁–C₈ alkyl,    -   C₁–C₈ alkoxy,    -   C₃–C₇ cycloalkyl,    -   C₁–C₄ haloalkyl,    -   (D)heterocyclyl    -   (D)C(O)R⁸,    -   (D)C(O)(CH₂)_(n)N(R⁸)₂,    -   C₁–C₈ alkyl-N(R⁸)₂,    -   (D))OR⁸,    -   (D)OCOR⁸,    -   (D)OC(O)N(R⁸)₂,    -   (D)N(R⁸)₂,    -   (D)NR⁸C(O)R⁸,    -   (D)NR⁸C(O)OR⁸,    -   (D)NR⁸C(O)N(R⁸)₂,    -   (D)NR⁸SO₂R⁸,    -   (D)SR⁸,    -   (D)SOR⁸,    -   (D)SO₂R⁸, or    -   (D)SO₂N(R⁸)₂;-   each R¹⁰ is independently:    -   hydrogen, (C₁–C₈)alkyl, C(O)C₁–C₈ alkyl, aryl or C₃–C₇        cycloalkyl;-   each R¹¹ is independently:    -   hydrogen,    -   C₁–C₈ alkyl,    -   (D)aryl,    -   (D)heteroaryl    -   (CH₂)_(n)N(R⁸)₂,    -   (CH₂)_(n)NR⁸C(O)C₁–C₄ alkyl,    -   (CH₂)_(n)NR⁸SO₂C₁–C₄ alkyl,    -   (CH₂)_(n)SO₂N(R⁸)₂,    -   (CH₂)_(n)[O]_(q)C₁–C₈ alkyl,    -   (CH₂)_(n)[O]_(q)(CH₂)_(n)NR⁸COR⁸,    -   (CH₂)_(n)[O]_(q)(CH₂)_(n)NR⁸SO₂R⁸,    -   (CH₂)_(n)[O]_(q)-heterocyclyl or    -   (CH₂)_(n)[O]_(q)(C₁–C₈ alkyl)-heterocyclyl; and    -   wherein n is 2–8;-   each R¹² is independently:    -   hydrogen,    -   C₁–C₈ alkyl,    -   (D)C₃–C₇ cycloalkyl,    -   (D)phenyl    -   C(O)C₁–C₈ allyl,    -   C(O)phenyl,    -   SO₂C₁–C₈ alkyl or    -   SO₂-phenyl;    -   D is a bond or —(CH₂)_(n)—;    -   n is 0–8;    -   p is 0–4;    -   q is 0–1;    -   r is 1–2; and    -   y is 0–1.

The compounds of the present invention are useful in preventing ortreating obesity or diabetes mellitus in a mammal comprising theadministration of a therapeutically effective amount of the compound offormula I.

The compounds of the present invention are also useful in preventing ortreating male or female sexual dysfunction in mammal, more specificallyerectile dysfunction, comprising the administration of a therapeuticallyeffective amount of the compound of formula I.

Also within the scope of the present invention is a pharmaceuticalcomposition or formulation which comprises a pharmaceutical carrier andat least one compound of formula I or its pharmaceutically acceptablesalts or stereoisomers thereof.

The present invention further includes a process of making apharmaceutical composition or formulation comprising a compound offormula I or its pharmaceutically acceptable salt or stereoisomersthereof and a pharmaceutically acceptable carrier.

The present invention further includes a process of preparing a compoundof formula I.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to melanocortin receptor agonists, andmore particularly piperazine and piperidine derivatives as melanocortinreceptor agonists. The compounds of present invention are useful for thetreatment or prevention of diseases and disorders responsive to theactivation of melanocortin receptors, such as obesity, diabetes andsexual dysfunction including erectile dysfunction and female sexualdysfunction.

An embodiment of the present invention is a compound of formula I:

or a pharmaceutically acceptable salts or stereoisomers thereof, wherein

-   G is CR¹ or N;-   A is C₁–C₈ alkyl or C₃–C₇ cycloalkyl;-   L and L¹ are independently: hydrogen or together oxo;-   T is:

-   R is: when y is 1;    -   N(R⁸)₂,    -   NR⁸COR⁸,    -   NR⁸CON(R⁸)₂,    -   NR⁸C(O)OR⁸,    -   NR⁸C(R⁸)═N(R⁸),    -   NR⁸SO₂R⁸ or    -   NR⁸SO₂N(R⁸)₂;-   R is: when y is 0 or 1;    -   heterocyclyl, provided that when y is 0, a heteroatom is not        directly connected to oxygen or adjacent to a carbon that        connected to oxygen; and    -   wherein the heterocyclyl contains at least one nitrogen in the        ring and is optionally substituted with one to five substituents        independently selected from R⁸;-   R¹ is independently:    -   hydrogen, CONH(C₁–C₈ alkyl), C₁–C₈ alkyl, (D)phenyl, (D)C₃–C₇        cycloalkyl or oxo, provided that oxo is not attached to the same        carbon that attached to nitrogen which forms an amide bond when        G is N;-   R² is independently:    -   hydrogen,    -   halo    -   C₁–C₈ alkyl,    -   C₁–C₈ alkylsulfonyl,    -   (D)C₃–C₇ cycloalkyl or    -   C₁–C₄ haloalkyl;-   R³ is independently: aryl or thienyl;    -   wherein aryl and thienyl are optionally substituted with one to        three substituents selected from the group consisting of:    -   cyano, halo, C₁–C₈ alkyl, (D)C₃–C₇ cycloalkyl, C₁–C₄ alkoxy,        C₁–C₄ haloalkyl and C₁–C₄ haloalkyloxy;-   R⁴ is independently:    -   hydrogen, C₁–C₈ alkyl, C(O)R⁸, C(O)OR⁸, C₃–C₇ cycloalkyl or        (CH₂)_(n)O(C₁–C₈ alkyl), wherein n is 2–8;-   each R⁸ is independently:    -   hydrogen,    -   phenyl    -   C₁–C₈ alkyl,    -   C₁–C₈ alkylsulfonyl,    -   C(O)C₁–C₈ alkyl,    -   C(O)aryl, wherein aryl being phenyl or naphthyl,    -   SO₂-aryl, wherein aryl being phenyl or naphthyl,    -   (D)C₃–C₇ cycloalkyl or    -   (CH₂)_(n)C₁–C₄ haloalkyl, wherein n is 1–8;-   each R⁹ is independently:    -   hydrogen,    -   hydroxy,    -   (D)cyano,    -   halo,    -   C₁–C₈ alkyl,    -   C₁–C₈ alkoxy,    -   C₃–C₇ cycloalkyl,    -   C₁–C₄ haloalkyl,    -   (D)heterocyclyl    -   (D)C(O)R⁸,    -   (D)C(O)(CH₂)_(n)N(R⁸)₂,    -   C₁–C₈ alkyl-N(R⁸)₂,    -   (D)OR⁸,    -   (D)OCOR⁸,    -   (D)OC(O)N(R⁸)₂,    -   (D)N(R⁸)₂,    -   (D)NR⁸C(O)R⁸,    -   (D)NR⁸C(O)OR⁸,    -   (D)NR⁸C(O)N(R⁸)₂,    -   (D)NR⁸SO₂R⁸,    -   (D)SR⁸,    -   (D)SOR⁸,    -   (D)SO₂R⁸, or    -   (D)SO₂N(R⁸)₂;-   each R¹⁰ is independently:    -   hydrogen, (C₁–C₈)alkyl, C(O)C₁–C₈ alkyl, aryl or C₃–C₇        cycloalkyl;-   each R¹¹ is independently:    -   hydrogen,    -   C₁–C₈ alkyl,    -   (D)aryl,    -   (D)heteroaryl    -   (CH₂)_(n)N(R⁸)₂,    -   (CH₂)_(n)NR⁸C(O)C₁–C₄ alkyl,    -   (CH₂)_(n)NR⁸SO₂C₁–C₄ alkyl,    -   (CH₂)_(n)SO₂N(R⁸)₂,    -   (CH₂)_(n)[O]_(q)C₁–C₈ alkyl,    -   (CH₂)_(n)[O]_(q)(CH₂)_(n)NR⁸COR⁸,    -   (CH₂)_(n)[O]_(q)(CH₂)_(n)NR⁸SO₂R⁸,    -   (CH₂)_(n)[O]_(q)-heterocyclyl or    -   (CH₂)_(n)[O]_(q)(C₁–C₈ alkyl)-heterocyclyl; and    -   wherein n is 2–8;-   each R¹² is independently:    -   hydrogen,    -   C₁–C₈ alkyl,    -   (D)C₃–C₇ cycloalkyl,    -   (D)phenyl    -   C(O)C₁–C₈ allyl,    -   C(O)phenyl,    -   SO₂C₁–C₈ alkyl or    -   SO₂-phenyl;-   D is a bond or —(CH₂)_(n)—;-   n is 0–8;-   p is 0–4;-   q is 0–1;-   r is 1–2; and-   y is 0–1.

The compound of the present invention as recited above, wherein theheterocyclyl is a 4-, 5- or 6-membered ring containing one nitrogenatom.

The compound of the present invention as recited above, wherein theheterocyclyl is a 6-membered ring containing one nitrogen and one oxygenatom.

The compound of the present invention as recited above, wherein R³ isphenyl optionally para-substituted with fluoro, chloro, bromo, iodo,benzyloxy, methoxy or methyl. The preferred R³ is phenylpara-substituted with chloro, fluoro or methoxy.

The compound of the present invention as recited above, wherein R⁴ ishydrogen.

The compound of the present invention as recited above, wherein—(CH₂)_(n)—T is:

* denotes a chiral carbon atom having a R or S configuration.

The compound of the present invention as recited above, wherein L and L¹are together oxo and the chiral carbon has R configuration.

A preferred embodiment of the present invention provides a compound offormula II,

or a pharmaceutically acceptable salts or stereoisomers thereof.

Yet another preferred embodiment of the present invention provides acompound of formula III,

or a pharmaceutically acceptable salts or stereoisomers thereof.

Yet another preferred embodiment of the present invention provides acompound of formula IV,

or a pharmaceutically acceptable salts or stereoisomers thereof.

Yet another preferred embodiment of the present invention provides acompound of formula V,

or a pharmaceutically acceptable salts or stereoisomers thereof.

Yet another preferred embodiment of the present invention provides acompound of formula VI,

or a pharmaceutically acceptable salts or stereoisomers thereof.

The substituents of the compound of present invention as recited abovein formula (II) to (VI) are as follows:

-   A is C₁–C₈ alkyl or C₃–C₇ cycloalkyl;-   r is 0 or 1;-   y is 0 or 1;-   D is a bond or —(CH₂)_(n)—;-   n is 0–8;-   R is: when y is 1;    -   N(R⁸)₂,    -   NR⁸COR⁸,    -   NR⁸CON(R⁸)₂,    -   NR⁸C(O)OR⁸,    -   NR⁸C(R⁸)═N(R⁸),    -   NR⁸SO₂R⁸ or    -   NR⁸SO₂N(R⁸)₂;-   R is: when y is 0 or 1;    -   heterocyclyl, provided that when y is 0, a heteroatom is not        directly connected to oxygen or adjacent to a carbon that        connected to oxygen; and    -   wherein the heterocyclyl contains at least one nitrogen in the        ring and are optionally substituted with one to five        substituents independently selected from R⁸;-   each R⁸ is independently:    -   hydrogen,    -   phenyl    -   C₁–C₈ alkyl,    -   C₁–C₈ alkylsulfonyl,    -   C(O)C₁–C₈ alkyl,    -   C(O)aryl, wherein aryl being phenyl or naphthyl,    -   SO₂-aryl, wherein aryl being phenyl or naphthyl,    -   (D)C₃–C₇ cycloalkyl or    -   (CH₂)_(n)C₁–C₄ haloalkyl, wherein n is 1–8;-   each R¹⁰ is independently:    -   hydrogen, (C₁–C₈)alkyl, C(O)C₁–C₈ alkyl, aryl or C₃–C₇        cycloalkyl.

The compound of present invention as recited above in formula (II)–(VI),wherein O—(A)_(y)—R is attached to ortho position of the phenyl ring.

The compound of present invention as recited above in formula (I)–(VI),wherein the heterocyclyl is a 4-, 5- or 6-membered ring containing onenitrogen atom.

The compound of present invention as recited above in formula (II)–(VI),wherein the nitrogen is substituted with one substituent selected fromR⁸ when y is 0.

The compound of present invention as recited above in formula (II)–(VI),wherein the heterocyclyl is a 6-membered ring containing one nitrogenand one oxygen atom.

The most preferred compounds of the present invention are:

Name of Compound Compound(N-(1-(4-R-chlorobenzyl)-2-{4-[2-(1-methyl-S-piperidin-3-yloxy)-phenyl]-piperazin-1-yl}-2-oxo-ethyl)-2-(2,3-dihydro-1H-isoindol-1-yl)-aceamide,trihydrochloride

(N-(1-(4-R-chlorobenzyl)-2-oxo-2-{4-[2-(R-piperidin-3-yloxy)-phenyl]-piperazin-1-yl}ethyl)-2-(2,3-dihydro-1H-isoindol-1-yl)-aceamide,trihydrochloride

2-(2,3-dihydro-1H-isoindol-1-yl)-N-(1-(4-methoxy-benzyl)-2-{4-[2-(1-methyl-piperidin-3-yloxy)-phenyl]-piperazin-1-yl}-2-oxo-ethyl)-acetamide,trihydrochloride

Also encompassed by the present invention is a pharmaceuticalcomposition or formulation, which comprises a pharmaceutical carrier andat least one compound of formula I or its pharmaceutically acceptablesalts or stereoisomers thereof. The pharmaceutical composition and orformulation may optionally further include a second active ingredientselected from the group consisting of an insulin sensitizer, insulinmimetic, sulfonylurea, alpha-glucosidase inhibitor, HMG-CoA reductaseinhibitor, sequestrant cholesterol lowering agent, beta 3 adrenergicreceptor agonist, neuropeptide Y antagonist, phosphodiester V inhibitor,and an alpha 2 adrenergic receptor antagonist.

Yet another aspect of the present invention is a process of making apharmaceutical composition comprising a compound of formula I or itspharmaceutically acceptable salt or stereoisomers thereof as recitedabove and a pharmaceutically acceptable carrier.

Yet another aspect of the present invention is a method of preventing ortreating obesity or diabetes mellitus in mammal comprising theadministration of a therapeutically effective amount of the compound offormula I.

Yet anther aspect of the present invention is a method of preventing ortreating male or female sexual dysfunction in mammal, more specificallythe male or female sexual dysfunction, comprising the administration ofa therapeutically effective amount of the compound of formula I.

Yet another aspect of the present invention is a process for preparing acompound of formula I:

or a pharmaceutically acceptable salts or stereoisomers thereof, wherein

-   G is CR¹ or N;-   A is C₁–C₈ alkyl or C₃–C₇ cycloalkyl;    -   —CLL¹—(CH₂)_(n)—T is:

wherein R₁ is hydrogen, C₁–C₈ alkyl, Boc, CBZ, FMOC, phenyl or (C₁–C₈alkyl)phenyl;

-   Q represents a moiety:

-   R is: when y is 1;    -   N(R⁸)₂,    -   NR⁸COR⁸,    -   NR⁸CON(R⁸)₂,    -   NR⁸C(O)OR⁸,    -   NR⁸C(R⁸)═N(R⁸),    -   NR⁸SO₂R⁸ or    -   NR⁸SO₂N(R⁸)₂;-   R is: when y is 0 or 1;    -   heterocyclyl, provided that when y is 0, a heteroatom is not        directly connected to oxygen or adjacent to a carbon that        connected to oxygen; and    -   wherein the heterocyclyl contains at least one nitrogen in the        ring and is optionally substituted with one to five substituents        independently selected from R⁸;-   R¹ is independently:    -   hydrogen, CONH(C₁–C₈ alkyl), C₁–C₈ alkyl, (D)phenyl, (D)C₃–C₇        cycloalkyl or oxo, provided that oxo is not attached to the same        carbon that attached to nitrogen which forms an amide bond when        G is N;-   R² is independently:    -   hydrogen,    -   halo    -   C₁–C₈ alkyl,    -   C₁–C₈ alkylsulfonyl,    -   (D)C₃–C₇ cycloalkyl or    -   C₁–C₄ haloalkyl;-   R³ is independently: aryl or thienyl;    -   wherein aryl and thienyl are optionally substituted with one to        three substituents selected from the group consisting of:    -   cyano, halo, C₁–C₈ alkyl, (D)C₃–C₇ cycloalkyl, C₁–C₄ alkoxy,        C₁–C₄ haloalkyl and C₁–C₄ haloalkyloxy;-   R⁴ is independently:    -   hydrogen, C₁–C₈ alkyl, C(O)R⁸, C(O)OR⁸, C₃–C₇ cycloalkyl or        (CH₂)_(n)O(C₁–C₈ alkyl), wherein n is 2–8;-   each R⁸ is independently:    -   hydrogen,    -   phenyl    -   C₁–C₈ alkyl,    -   C₁–C₈ alkylsulfonyl,    -   C(O)C₁–C₈ alkyl,    -   C(O)aryl, wherein aryl being phenyl or naphthyl,    -   SO₂-aryl, wherein aryl being phenyl or naphthyl,    -   (D)C₃–C₇ cycloalkyl or    -   (CH₂)_(n)C₁–C₄ haloalkyl, wherein n is 1–8;-   each R⁹ is independently:    -   hydrogen,    -   hydroxy,    -   (D)cyano,    -   halo,    -   C₁–C₈ alkyl,    -   C₁–C₈ alkoxy,    -   C₃–C₇ cycloalkyl,    -   C₁–C₄ haloalkyl,    -   (D)heterocyclyl    -   (D)C(O)R⁸,    -   (D)C(O)(CH₂)_(n)N(R⁸)₂,    -   C₁–C₈ alkyl-N(R⁸)₂,    -   (D)OR⁸,    -   (D)OCOR⁸,    -   (D)OC(O)N(R⁸)₂,    -   (D)N(R⁸)₂,    -   (D)NR⁸C(O)R⁸,    -   (D)NR⁸C(O)OR⁸,    -   (D)NR⁸C(O)N(R⁸)₂,    -   (D)NR⁸SO₂R⁸,    -   (D)SR⁸,    -   (D)SOR⁸,    -   (D)SO₂R⁸, or    -   (D)SO₂N(R⁸)₂;-   each R¹⁰ is independently:    -   hydrogen, (C₁–C₈)alkyl, C(O)C₁–C₈ alkyl, aryl or C₃–C₇        cycloalkyl;-   D is a bond or —(CH₂)_(n)—;-   n is 0–8;-   p is 0–4;-   q is 0–1;-   r is 1–2; and-   y is 0–1.    comprising the steps of:

a) reacting a compound having a structural formula 1,

with CH₂CH═C(O)OR^(a) wherein R^(a) is hydrogen or C₁–C₈ alkyl and X ishalo, in the presence of a catalyst and a base in a suitable organicsolvent to give the compound of formula 2,

b) reductively aminating the compound of formula 2 in the presence ofamine in an acidic condition to give a compound of formula 3,

c) cyclizing the compound of formula 3 by Michael addition to give acompound of formula 4 or stereoisomers thereof,

d) coupling the compound of formula 4 or stereoisomers thereof, whereinR^(a) of compound 4 is H, with a compound of formula 5,

wherein R^(a) of compound 5 is C₁–C₈ alkyl, to give a compound offormula 6;

e) coupling the compound of formula 6, wherein R^(a) is H, with acompound having a structural,

to afford the compound of formula 1.

The process of present invention as recited above, wherein

in Step (a) is 2-boromobenzaldehydes.

The process of present invention as recited above, wherein CH₂CH═C(O)ORin Step (a) is methylacrylate.

The process of present invention as recited above, wherein the catalystin Step (a) is selected from the group consisting of: Pd(Ph₃P)₂Cl₂,Pd(Ph₃P)₄Cl₂, Pd(Ph₃P)₄, Pd(Ph₃P)₂Cl₂/CuI, Pd(OAc)₂/Ph₃P—Bu₄NBr,Pd(Ph₃P)₄Cl₂/H₂ and Pd(OAc)₂/P(O-tol)₃; and wherein the base in Step (a)is NR₃ wherein R is hydrogen or C₁–C₈ alkyl.

The process of present invention as recited above, wherein the amine inStep (b) is selected from the group consisting of: benzylamine,alpha-methylbenzylamine and BocNH₂.

The process of present invention as recited above, wherein the Step (b)further comprises reducing of intermediate imine compound in thepresence of reducing agent, the reducing agent being selected from thegroup consisting of: NaCNBH₃, Na(OAc)₃BH, NaBH₄/H+, and a combination ofEt₃SiH and TFA in CH₃CN or CH₂Cl₂.

The process of present invention as recited above, wherein thestereoisomer of compound of formula 4 in Step (c) is a compound offormula 4a.

The process of present invention as recited above, wherein the compoundof formula 4a is prepared by asymmetric hydrogenation of a compoundhaving structural formula,

The process of present invention as recited above, wherein the Michaeladdition in Step (c) is carried out in a basic workup condition.

The process of present invention as recited above, wherein the Step (e)further comprises deprotecting or protecting of the compound of formula(4) at NR₁.

Yet another aspect of the present invention is a process for preparing acompound of formula I:

or a pharmaceutically acceptable salts or stereoisomers thereof, wherein

-   G is CR¹ or N;-   A is C₁–C₈ alkyl or C₃–C₇ cycloalkyl;    -   —CLL¹—(CH₂)_(n)—T is:

-   Q represents a moiety:

-   R is: when y is 1;    -   N(R⁸)₂,    -   NR⁸COR⁸,    -   NR⁸CON(R⁸)₂,    -   NR⁸C(O)OR⁸,    -   NR⁸C(R⁸)═N(R⁸),    -   NR⁸SO₂R⁸ or    -   NR⁸SO₂N(R⁸)₂;-   R is: when y is 0 or 1;    -   heterocyclyl, provided that when y is 0, a heteroatom is not        directly connected to oxygen or adjacent to a carbon that        connected to oxygen; and    -   wherein the heterocyclyl contains at least one nitrogen in the        ring and is optionally substituted with one to five substituents        independently selected from R⁸;-   R¹ is independently:    -   hydrogen, CONH(C₁–C₈ alkyl), C₁–C₈ alkyl, (D)phenyl, (D)C₃–C₇        cycloalkyl or oxo, provided that oxo is not attached to the same        carbon that attached to nitrogen which forms an amide bond when        G is N;-   R² is independently:    -   hydrogen,    -   halo    -   C₁–C₈ alkyl,    -   C₁–C₈ alkylsulfonyl,    -   (D)C₃–C₇ cycloalkyl or    -   C₁–C₄ haloalkyl;-   R³ is independently: aryl or thienyl;    -   wherein aryl and thienyl are optionally substituted with one to        three substituents selected from the group consisting of:    -   cyano, halo, C₁–C₈ alkyl, (D)C₃–C₇ cycloalkyl, C₁–C₄ alkoxy,        C₁–C₄ haloalkyl and C₁–C₄ haloalkyloxy;-   R⁴ is independently:    -   hydrogen, C₁–C₈ alkyl, C(O)R⁸, C(O)OR⁸, C₃–C₇ cycloalkyl or        (CH₂)_(n)O(C₁–C₈ alkyl), wherein n is 2–8;-   each R⁸ is independently:    -   hydrogen,    -   phenyl    -   C₁–C₈ alkyl,    -   C₁–C₈ alkylsulfonyl,    -   C(O)C₁–C₈ alkyl,    -   C(O)aryl, wherein aryl being phenyl or naphthyl,    -   SO₂-aryl, wherein aryl being phenyl or naphthyl,    -   (D)C₃–C₇ cycloalkyl or    -   (CH₂)_(n)C₁–C₄ haloalkyl, wherein n is 1–8;-   each R⁹ is independently:    -   hydrogen,    -   hydroxy,    -   (D)cyano,    -   halo,    -   C₁–C₈ alkyl,    -   C₁–C₈ alkoxy,    -   C₃–C₇ cycloalkyl,    -   C₁–C₄ haloalkyl,    -   (D)heterocyclyl    -   (D)C(O)R⁸,    -   (D)C(O)(CH₂)_(n)N(R⁸)₂,    -   C₁–C₈ alkyl-N(R⁸)₂,    -   (D))OR⁸,    -   (D)OCOR⁸,    -   (D)OC(O)N(R⁸)₂,    -   (D)N(R⁸)₂,    -   (D)NR⁸C(O)R⁸,    -   (D)NR⁸C(O)OR⁸,    -   (D)NR⁸C(O)N(R⁸)₂,    -   (D)NR⁸SO₂R⁸,    -   (D)SR⁸,    -   (D)SOR⁸,    -   (D)SO₂R⁸, or    -   (D)SO₂N(R⁸)₂;-   each R¹¹ is independently: hydrogen, (C₁–C₈)alkyl,-   D is a bond or —(CH₂)_(n)—;-   n is 0–8;-   p is 0–4;-   q is 0–1;-   r is 1–2; and-   y is 0–1.    comprising the steps of:

a) esterifying a compound of formula 1,

with an alcohol R^(a)OH to form a compound of formula 2,

wherein R^(a) is C₁–C₄ alkyl or (D)phenyl;

b) reacting a compound of formula 2 with R¹¹COR¹¹ to form a compound offormula 3,

wherein R¹¹ is independently hydrogen or C₁–C₄ alkyl;

c) reacting a compound of formula 3 with an activating group to form acompound of formula 4,

wherein A is an activating group;

d) deoxygenating the compound of formula 4 by hydrogenation to afford acompound of formula 5,

e) optionally reacting the compound of formula 5 with an inorganic baseto form a compound of formula 6,

wherein HA is an acidic and M is a univalent cation;

-   -   f) resolving the compound of formula 5 or formula 6 to afford a        chiral compound of formula 7,

wherein M is hydrogen and R^(a) is H or R^(a);

g) coupling the compound of formula 7 with a compound of formula 8,

to afford a compound of formula 9,

h) coupling the compound of formula 9 with a compound having a formula,

to afford a compound of formula I.

Yet another aspect of the present invention is a process for preparing acompound of formula I,

or a pharmaceutically acceptable salts or stereoisomers thereof, wherein

-   G is CR¹ or N;-   A is C₁–C₈ alkyl or C₃–C₇ cycloalkyl;    -   —CLL¹—(CH₂)_(n)—T is:

-   Q represents a moiety:

-   R is: when y is 1;    -   N(R⁸)₂,    -   NR⁸COR⁸,    -   NR⁸CON(R⁸)₂,    -   NR⁸C(O)OR⁸,    -   NR⁸C(R⁸)═N(R⁸),    -   NR⁸SO₂R⁸ or    -   NR⁸SO₂N(R⁸)₂;-   R is: when y is 0 or 1;    -   heterocyclyl, provided that when y is 0, a heteroatom is not        directly connected to oxygen or adjacent to a carbon that        connected to oxygen; and    -   wherein the heterocyclyl contains at least one nitrogen in the        ring and is optionally substituted with one to five substituents        independently selected from R⁸;-   R¹ is independently:    -   hydrogen, CONH(C₁–C₈ alkyl), C₁–C₈ alkyl, (D)phenyl, (D)C₃–C₇        cycloalkyl or oxo, provided that oxo is not attached to the same        carbon that attached to nitrogen which forms an amide bond when        G is N;-   R² is independently:    -   hydrogen,    -   halo    -   C₁–C₈ alkyl,    -   C₁–C₈ alkylsulfonyl,    -   (D)C₃–C₇ cycloalkyl or    -   C₁–C₄ haloalkyl;-   R³ is independently: aryl or thienyl;    -   wherein aryl and thienyl are optionally substituted with one to        three substituents selected from the group consisting of:    -   cyano, halo, C₁–C₈ alkyl, (D)C₃–C₇ cycloalkyl, C₁–C₄ alkoxy,        C₁–C₄ haloalkyl and C₁–C₄ haloalkyloxy;-   R⁴ is independently:    -   hydrogen, C₁–C₈ alkyl, C(O)R⁸, C(O)OR⁸, C₃–C₇ cycloalkyl or        (CH₂)_(n)O(C₁–C₈ alkyl), wherein n is 2–8;-   each R⁸ is independently:    -   hydrogen,    -   phenyl    -   C₁–C₈ alkyl,    -   C₁–C₈ alkylsulfonyl,    -   C(O)C₁–C₈ alkyl,    -   C(O)aryl, wherein aryl being phenyl or naphthyl,    -   SO₂-aryl, wherein aryl being phenyl or naphthyl,    -   (D)C₃–C₇ cycloalkyl or    -   (CH₂)_(n)C₁–C₄ haloalkyl, wherein n is 1–8;-   each R⁹ is independently:    -   hydrogen,    -   hydroxy,    -   (D)cyano,    -   halo,    -   C₁–C₈ alkyl,    -   C₁–C₈ alkoxy,    -   C₃–C₇ cycloalkyl,    -   C₁–C₄ haloalkyl,    -   (D)heterocyclyl    -   (D)C(O)R⁸,    -   (D)C(O)(CH₂)_(n)N(R⁸)₂,    -   C₁–C₈ alkyl-N(R⁸)₂,    -   (D)OR⁸,    -   (D)OCOR⁸,    -   (D)OC(O)N(R⁸)₂,    -   (D)N(R⁸)₂,    -   (D)NR⁸C(O)R⁸,    -   (D)NR⁸C(O)OR⁸,    -   (D)NR⁸C(O)N(R⁸)₂,    -   (D)NR⁸SO₂R⁸,    -   (D)SR⁸,    -   (D)SOR⁸,    -   (D)SO₂R⁸, or    -   (D)SO₂N(R⁸)₂;-   each R¹⁰ is independently:    -   hydrogen, (C₁–C₈)alkyl, C(O)C₁–C₈ alkyl, aryl or C₃–C₇        cycloalkyl, or protecting group selected from Boc, CBZ or FMOC;-   each R¹¹ is independently: hydrogen or (C₁–C₈)alkyl;-   D is a bond or —(CH₂)_(n)—;-   n is 0–8;-   p is 0–4;-   q is 0–1;-   r is 1–2; and-   y is 0–1.    comprising the steps of:

a) reacting a compound formula 1:

wherein X is halo, and R¹¹ is independently, hydrogen or C1–C4 alkyl,with CNCH₂CO₂R^(a) wherein R^(a) is C₁–C₈ alkyl or benzyl to afford acompound of formula 2:

b) protecting the compound of formula 2 to form the compound of formula3:

c) hydrogenating the compound of formula 3 to afford a compound offormula 4:

d) coupling the compound of formula 4 wherein R^(a′) is hydrogen orR^(a), with a compound of formula 5,

to afford a compound of formula 6,

e) coupling the compound of formula 6 with a compound having a formula,

to afford a compound of formula I.

Throughout the instant application, the following terms have theindicated meanings:

The term “alkyl,” unless otherwise indicated, refers to those alkylgroups of a designated number of carbon atoms of either a straight orbranched saturated configuration. Examples of “alkyl” includes, but arenot limited to methyl, ethyl, n-propyl. isopropyl, n-butyl, isobutyl,sec-butyl and t-butyl, pentyl, hexyl, neopenyl, isopentyl and the like.

The term “alkenyl” means hydrocarbon chain of a specified number ofcarbon atoms of either a straight or branched configuration and havingat least one carbon-carbon double bond, which may occur at any pointalong the chain, such as ethenyl, propenyl, butenyl, pentenyl, vinyl,alkyl, 2-butenyl and the like.

The term “haloalkyl” is an alkyl group of indicated number of carbonatoms, which is substituted with one to five halo atoms selected from F,Br, Cl, and I. An example of a haloalkyl group is trifluoromethyl.

The term “alkoxy” represents an alkyl group of indicated number ofcarbon atoms attached through an oxygen bridge, such as methoxy, ethoxy,propoxy, isopropoxy, butoxy, tert-butoxy, pentoxy, and the like.

The term “cycloalkyl” refers to a ring composed of 3 to 7 methylenegroups, each of which may be optionally substituted with otherhydrocarbon substituents. Examples of cycloalkyl includes, but are notlimited to: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl andcycloheptyl, and the like.

The term “halo” refers to fluoro, chloro, bromo and iodo.

The term “haloalkyloxy” represents a haloalkyl group of indicated numberof carbon atoms attached through an oxygen bridge, such as OCF₃.

The term “aryl” refers to phenyl, naphthyl, anthracenyl, phenanthrenyland the like.

The term “heteroaryl” refers to monocyclic or bicyclic aromatic ring of5- to 10-carbon atoms containing from one to four heteroatoms selectedfrom O, N, or S. Examples of heteroaryl are, but are not limited tofuranyl, thienyl, thiazolyl, imidazolyl, isoxazoyl, oxazoyl, pyrazoyl,pyrrolyl, pyrazinyl, pyridyl, pyrimidyl, and purinyl, cinnolinyl,benzothienyl, benzotriazolyl, benzoxazolyl, quinoline, isoquinoline andthe like.

The “heterocyclyl” is defined as a monocyclic, bicyclic, or tricyclicring of 5 to 14 carbon atoms which are saturated or partially saturatedcontaining from one to four heteroatoms selected from N, O or S. The“heterocyclyl” includes “nitrogen containing heterocyclyl,” whichcontains from one to four nitrogen atoms and optionally further containsone other heteroatom selected from O or S.

A mammal as used in here includes a human and a warm-blooded animal suchas a cat, a dog and the like.

The term “composition” or “formulation”, as in pharmaceuticalcomposition or formulation, is intended to encompass a productcomprising the active ingredient(s), and the inert ingredient(s) thatmake up the carrier. Accordingly, the pharmaceutical compositions of thepresent invention encompass any composition made by admixing a compoundof the present invention (a compound of formula 1) and apharmaceutically acceptable carrier.

The term “pharmaceutical” when used herein as an adjective meanssubstantially non-deleterious to the recipient mammal.

The term “unit dosage form” refers to physically discrete units suitableas unitary dosages for human subjects and other non-human animals suchas warm-blooded animals each unit containing a predetermined quantity ofactive ingredient (a compound of formula I) calculated to produce thedesired therapeutic effect in association with a suitable pharmaceuticalcarrier.

The term “treating” or “preventing” as used herein includes itsgenerally accepted meanings, i.e., preventing, prohibiting, restraining,alleviating, ameliorating, slowing, stopping, or reversing theprogression or severity of a pathological condition, or sequela thereofas described herein.

“Erectile dysfunction” is a disorder involving the failure of a malemammal to achieve erection, ejaculation, or both. Symptoms of erectiledysfunction include an inability to achieve or maintain an erection,ejaculatory failure, premature ejaculation, inability to achieve anorgasm. An increase in erectile dysfunction is often associated with ageand is generally caused by a physical disease or as a side effect ofdrug treatment.

“Female sexual dysfunction” encompasses, without limitation, conditionssuch as a lack of sexual desire and related arousal disorders, inhibitedorgasm, lubrication difficulties, and vaginismus.

Because certain compounds of the invention contain an acidic moiety(e.g., carboxy), the compound of formula I may exist as a pharmaceuticalbase addition salt thereof. Such salts include those derived frominorganic bases such as ammonium and alkali and alkaline earth metalhydroxides, carbonates, bicarbonates and the like, as well as saltsderived from basic organic amines such as aliphatic and aromatic amines,aliphatic diamines, hydroxy alkamines, and the like.

Because certain compounds of the invention contain a basic moiety (e.g.,amino), the compound of formula I can also exist as a pharmaceuticalacid addition salt. Such salts include sulfate, pyrosulfate, bisulfate,sulfite, bisulfite, phosphate, mono-hydrogenphosphate,dihydrogenphosphate, metaphosphate, pyrophosphate, chloride, bromide,iodide, acetate, propionate, decanoate, caprylate, acrylate, formate,isobutyrate, heptanoate, propiolate, oxalate, malonate, succinate,suberate, sebacate, fumarate, maleate, 2-butyne-1,4 dioate,3-hexyne-2,5-dioate, benzoate, chlorobenzoate, hydroxybenzoate,methoxybenzoate, phthalate, xylenesulfonate, phenylacetate,phenylpropionate, phenylbutyrate, citrate, lactate, hippurate,beta-hydroxybutyrate, glycollate, maleate, tartrate, methanesulfonate,propanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate,mandelate and the like salts. Preferred salt form of compound of formulaI is an acid addition salts, more specifically hydrochloride salt.

Some of the compounds described herein may exist as tautomers such asketoenol tautomers. The individual tautomers as well as mixtures thereofare encompassed within the scope of the present invention.

Utility

Compounds of formula I are effective as melanocortin receptormodulators, particularly as agonists of the human MC-4 receptor. Asmelanocortin receptor agonists, the compounds of formula I are useful inthe treatment of diseases, disorders or conditions responsive to theactivation of one or more of the melanocortin receptors including, butnot limited to, MC-1, MC-2, MC-3, MC4, and MC-5. Diseases, disorders orconditions receptive to treatment with a MC-4 agonist include thosementioned above and those described in WO 00/74679, the teachings ofwhich are herein incorporated by reference. In particular diseases,disorders or conditions receptive to treatment with a MC-4 agonistinclude obesity or diabetes mellitus, male or female sexual dysfunction,more specifically erectile dysfunction.

When describing various aspects of the present compounds of formula I,the terms “A domain”, “B domain” and “C domain” are used below. Thisdomain concept is illustrated below:

The following listing provides some of examples “A domain”, “B domain”and “C domain” of the compound of formula I. These listings are providedas illustrative purposes and as such are not meant to be limiting.

Examples of A Domain:

Examples of B Domain:

Examples of C Domain:

Formulation

The compound of formula I is preferably formulated in a unit dosage formprior to administration. Accordingly the present invention also includesa pharmaceutical composition comprising a compound of formula I and asuitable pharmaceutical carrier.

The present pharmaceutical compositions are prepared by known proceduresusing well-known and readily available ingredients. In making theformulations of the present invention, the active ingredient (a compoundof formula I) is usually mixed with a carrier, or diluted by a carrier,or enclosed within a carrier, which may be in the form of a capsule,sachet, paper or other container. When the carrier serves as a diluents,it may be a solid, semisolid or liquid material which acts as a vehicle,excipient or medium for the active ingredient. Thus, the compositionscan be in the form of tablets, pills, powders, lozenges, sachets,cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosol (asa solid or in a liquid medium), soft and hard gelatin capsules,suppositories, sterile injectable solutions and sterile packagedpowders.

Some examples of suitable carriers, excipients, and diluents includelactose, dextrose, sucrose, sorbitol, mannitol, starches, gum acacia,calcium phosphate, alginates, tragacanth, gelatin, calcium silicate,microcrystalline cellulose, polyvinylpyrrolidone, cellulose, watersyrup, methyl cellulose, methyl and propylhydroxybenzoates, talc,magnesium stearate and mineral oil. The formulations can additionallyinclude lubricating agents, wetting agents, emulsifying and suspendingagents, preserving agents, sweetening agents or flavoring agents. Thecompositions of the invention may be formulated so as to provide quick,sustained or delayed release of the active ingredient afteradministration to the patient.

Dosage:

The specific dose administered is determined by the particularcircumstances surrounding each situation. These circumstances include,the route of administration, the prior medical history of the recipient,the pathological condition or symptom being treated, the severity of thecondition/symptom being treated, and the age and sex of the recipient.Additionally, it would be understood that the therapeutic dosageadministered can be determined by the physician in the light of therelevant circumstances.

Generally, an effective minimum daily dose of a compound of formula I isabout 1, 5, 10, 15, or 20 mg. Typically, an effective maximum dose isabout 500, 100, 60, 50, or 40 mg. The suitable dose may be determined inaccordance with the standard practice in the medical arts of “dosetitrating” the recipient, which involves administering a low dose of thecompound initially and then gradually increasing the does until thedesired therapeutic effect is observed.

Route of Administration

The compounds may be administered by a variety of routes including theoral, rectal, transdermal, subcutaneous, topical, intravenous,intramuscular or intranasal routes.

Combination Therapy

Compounds of formula I may be used in combination with other drugs thatare used in the treatment of the diseases or conditions for whichcompounds of formula I are useful. Such other drugs may be administeredby a route and in an amount commonly used therefor, contemporaneously orsequentially with a compound of formula I. When a compound of formula Iis used contemporaneously with one or more other drugs, a pharmaceuticalcomposition containing such other drugs in addition to the compound offormula I is preferred. Accordingly, the pharmaceutical compositions ofthe present invention include those that also contain one or more otheractive ingredients in addition to a compound of formula I. Examples ofother active ingredients that may be combined with a compound of formulaI, either administered separately or in the same pharmaceuticalcompositions, include but are not limited to:

-   -   (a) insulin sensitizers including (i) PPARγ agonists such as the        glitazones (e.g. troglitazone, pioglitazone, englitazone,        MCC-555, BRL49653 and the like) and compounds disclosed in        WO97/27857, WO 97/28115, WO 97/28137 and WO97/27847; (ii)        biguanides such as metformin and phenformin;    -   (b) insulin or insulin mimetics;    -   (c) sulfonylureas such as tolbutamide and glipizide;    -   (d) α-glucosidase inhibitors (such as acarbose),    -   (e) cholesterol lowering agents such as (i) HMG-CoA reductase        inhibitors (lovastatin, simvastatin and pravastatin,        fluvastatin, atorvastatin, and other statins), (ii) sequestrants        (cholestyramine, colestipol and a dialkylaminoalkyl derivatives        of a cross-linked dextran), (iii) nicotinyl alcohol nicotinic        acid or a salt thereof, (iv) proliferator-activater receptor        α-agonists such as fenofibric acid derivatives (gemfibrozil,        clofibrat, fenofibrate and benzafibrate), (v) inhibitors of        cholesterol absorption such as β-sitosterol and acyl        CoA:cholesterol acyltransferase inhibitors such as        melinamide, (vi) probucol, (vii) vitamin E, and (viii)        thyromimetics;    -   (f) PPARδ agonists such as those disclosed in WO97/28149;    -   (g) antiobesity compounds such as fenfluramine, dexfenfluramine,        phentermine, sibutramine, orlistat, and β-3 adrenergic receptor        agonists;    -   (h) feeding behavior modifying agents such as neuropeptide Y        antagonists (e.g. neuropeptide Y5) as disclosed in WO 97/19682,        WO 97/20820, WO 97/20821, WO 97/20822 and WO 97/20823;    -   (i) PPARα agonists as described in WO 97/36579;    -   (j) PPARγ antagonists as described in WO97/10813;    -   (k) serotonin reuptake inhibitors, such as fluoxetine and        sertraline;    -   (l) growth hormone secretagogues such as MK-0677; and    -   (m) agents useful in the treatment of male and/or female sexual        dysfunction, such as phosphodiester V inhibitors including        sildenafil and ICI-351, and α-2 adrenergic receptor antagonists        including phentolamine mesylate; and dopamine-receptor agonists,        such as apomorphine.        Biological Assays        A. Binding Assay:

The radioligand binding assay is used to identify competitive inhibitorsof ¹²⁵I-NDP-α-MSH binding to cloned human MCRs using membranes fromstably transfected human embryonic kidney (HEK) 293 cells.

HEK 293 cells transfected with human or rat melanocortinin receptors aregrown either as adherent monolayers or suspension culture. Monolayercells are grown in roller bottle cultures at 37° C. and 5% CO₂/airatmosphere in a 3:1 mixture of Dulbecco's modified Eagle medium (DMEM)and Ham's F12 containing 25 mM L-glucose, 100 units/ml penicillin G, 100microgram/ml streptomyocin, 250 nanogram/ml amphoterin B, 300microgram/ml genticin and supplemented with 5% fetal bovine serum.Monolayer cells are adapted to suspension culture (Berg et al.,Biotechniques Vol. 14, No. 6, 1993) and are grown in either spinner orshaker flasks (37° C. and 7.5% CO₂/air overlay) in a modified DME/F12medium containing 0.1 mM CaCl₂, 2% equine serum and 100 microgram/mlsodium heparin to prevent cell-cell aggregation. Cells are harvested bycentrifugation, washed in PBS, and pellets are stored frozen at −80° C.until membrane preparations.

The cell pellets are resuspended in 10 volumes of membrane preparationbuffer (i.e., 1 g pellet to 10 ml buffer) having the followingcomposition: 50 mM Tris pH 7.5 @4° C., 250 mM sucrose, 1 mM MgCl₂,Complete® EDTA-free protease inhibitor tablet (Boehringer Mannheim), and24 micrograms/ml DNase I (Sigma, St. Louis, Mo.). The cells arehomogenized with a motor-driven dounce using 20 strokes, and thehomogenate is centrifuged at 38,000×g at 4° C. for 40 minutes. Thepellets are resuspended in membrane preparation buffer at aconcentration of 2.5–7.5 mg/ml and 1 milliliter aliquots of membranehomogenates are quickly frozen in liquid nitrogen and then stored at−80° C.

Solutions of a compound of formula I (300 picomolar to 30 micromolar) orunlabelled NDP-α-MSH (1 picomolar to 100 nanomolar) are added to 150microliters of membrane binding buffer to yield final concentrations(listed in parantheses). The membrane binding buffer has the followingcomposition: 25 mM HEPES pH 7.5; 10 mM CaCl₂; 0.3% BSA). One hundredfifty microliters of membrane binding buffer containing 0.5–5.0microgram membrane protein is added, followed by 50 nanomolar¹²⁵I-NDP-α-MSH to final concentration of 100 picomolar. Additionally,fifty microliters of SPA beads (5 mg/ml) are added and the resultingmixture is agitated briefly and incubated for 10 hours at r.t. Theradioactivity is quantified in a Wallac Trilux Microplate Scintillationcounter. IC₅₀ values obtained in competition assays are converted toaffinity constants (K_(i) values) using the Cheng-Prusoff equation:K_(i)=IC₅₀(1+D/K_(d)).

B. Functional Assay:

Functional cell based assays are developed to discriminate agonists andantagonists.

Agonist Assay: HEK 293 cells stably expressing a human melanocortinreceptor (see e.g., Yang, et al., Mol-Endocrinol., 11(3): 274–80, 1997)are dissociated from tissue culture flasks using a trypsin/EDTAsolution(0.25%; Life Technologies, Rockville, Md.). Cells are collectedby centrifugation and resuspended in DMEM (Life Technologies, Rockville,Md.) supplemented with 1% L-glutamine and 0.5% fetal bovine serum. Cellsare counted and diluted to 4.5×10⁵/ml.

A compound of formula I is diluted in dimethylsulfoxide (DMSO) (3×10⁻⁵to 3×10⁻¹⁰ M final concentrations) and 0.05 volume of compound solutionis added to 0.95 volumes of cell suspension; the final DMSOconcentration is 0.5%. After incubation at 37° C./5% CO₂ for 5 hours,cells are lysed by addition of luciferin solution (50 mM Tris, 1 mMMgCl₂, 0.2% Triton-X100, 5 mM DTT, 500 micromolar Coenzyme A, 150micromolar ATP, and 440 micromolar luciferin) to quantify the activityof the reporter gene luciferase, an indirect measurement ofintracellular cAMP production.

Luciferase activity is measured from the cell lysate using a WallacVictor 2 luminometer. The amount of lumen production which results froma compound of formula I is compared to that amount of lumens produced inresponse to NDP-α-MSH, defined as a 100% agonist, to obtain the relativeefficacy of a compound. The EC₅₀ is defined as the compoundconcentration that results in half maximal stimulation, when compared toits own maximal level of stimulation.

Antagonist assay: Antagonist activity is defined as the ability of acompound to block lumen production in response to NDP-α-MSH.Concentration-response curves are generated for NDP-α-MSH in the absenceand presence of a fixed concentration of a solution of a compound offormula I (10×K_(i) from binding assays). Suspensions of MCR-expressingcells are prepared and are incubated with NDP-α-MSH and compoundsolutions for 5 hours as described above. The assay is terminated by theaddition of luciferin reagent and lumen production is quantified.Antagonist potency is determined from the rightward shift of the EC₅₀value in the absence of a compound of formula I using the equation:K_(b)=Concentration of Antagonist/[(EC₅₀′/EC₅₀)−b 1].

Whole Cell cAMP Accumulation Assay

Compound Preparation

In the agonist assay, compounds are prepared as 10 M, and NDP-alpha-MSH(control) as 33.3 μM stock solutions in 100% DMSO. These are seriallydiluted in 100% DMSO. The compound plate is further diluted 1:200 incompound dilution buffer (HBSS-092, 1 mM Ascorbic Acid, 1 mM IBMX, 0.6%DMSO, 0.1% BSA). The final concentration range being 10 μM–100 pM forcompound and 33.33 nM–0.3 pM for control in 0.5% DMSO. Transfer 20 μlfrom this plate into four PET 96-well plates (all assays are performedin duplicate for each receptor).

Cell Culture and Cell Stimulation

HEK 293 cells stably transfected with the MC3R and MC4R were grown inDMEM containing 10% FBS and 1% Antibiotic/Antimycotic Solution. On theday of the assay the cells were dislodged with enzyme free celldissociation solution and resuspended in cell buffer (HBSS-092, 0.1%BSA, 10 mM HEPES) at 1×e6 cells/ml. Add 40 μl of cells/well to the PET96-well plates containing 20 microliter diluted compound and control.Incubate@37° C. in a water bath for 20 minutes. Stop the assay by adding50 μQuench Buffer (50 mM Na Acetate, 0.25% Triton X-100).

Radioligand Binding Assays

Radioligand binding assays were run in SPA buffer (50 mM Sodium Acetate,0.1% BSA). The beads, antibody and radioligand were diluted in SPAbuffer to provide sufficient volume for each 96-well plate. To eachquenched assay well was added 100 microliter cocktail containing 33.33microliter of beads, 33.33 microliter antibody and 33.33 microliter¹²⁵I-cAMP. This was based on a final concentration of 6.3 mg/ml beads,0.65% anti-goat antibody and 61 pM of ¹²⁵I-cAMP (containing 25000–30000CPM) in a final assay volume of 210 microliter. The plates were countedin a Wallac MicroBeta counter after a 12-hour incubation.

The data was converted to pmoles cAMP using a standard curve assayedunder the same conditions. The data was analyzed using Activity Basesoftware to generate agonist potencies (EC50) and percent relativeefficacy data to NDP-alpha-MSH.

C. In vivo Food Intake Models:

1) Daily food intake. Male Long-Evans rats are injectedintracerebroventricularly (ICV) with a test compound in 5 microliters of50% propylene glyco/artificial cerebrospinal fluid one hour prior toonset of dark cycle (12 hours). Food intake is determined by subtractingthe food weight remaining after 24 hours from food weight just prior toICV injection.

2) Acute Calorimetry. Male Long-Evans rats are administered testcompound by subcutaneous injection, intramuscular injection, intravenousinjection, intraperitoneal injection, ICV injection or by oral gavagebetween 0 and 5 hours after the onset of the dark cycle. Rats are placedinto a calorimetry chamber and the volume of oxygen consumed and volumeor carbon dioxide exhaled are measured each hour for 24 hours. Foodintake is measured for the 24 hour period as described in C. 1).Locomoter activity is measured when the rat breaks a series of infraredlaser beams when in the calorimeter. These measurements permitcalculation of energy expenditure, respiratory quotient and energybalance.

3) Food intake in diet induced obese mice. Male C57/B16J mice maintainedon a high fat diet (60% fat calories) for 6.5 months from 4 weeks of ageare dosed intraperitoneally with a compound of formula I. Food intakeand body weight are measured over an eight day period. Biochemicalparameters relating to obesity, including leptin, insulin, triglyceride,free fatty acid, cholesterol and serum glucose levels are determined.

D. Rat Ex Copula Assay:

Sexually mature male Caesarian Derived Sprague Dawley (CD) rats (over 60days old) are used with the suspensory ligament surgically removed toprevent retraction of the penis back into the penile sheath during theex copula evaluations. Animals receive food and water ad lib and arekept on a normal light/dark cycle. Studies are conducted during thelight cycle.

1) Conditioning to Supine Restraint for Ex Copula Reflex Tests. Thisconditioning takes about 4 days. Day 1, the animals are placed in adarkened restrainer and left for 15–30 minutes. Day 2, the animals arerestrained in a supine position in the restrainer for 15–30 minutes. Day3, the animals are restrained in the supine position with the penilesheath retracted for 15–30 minutes. Day 4, the animals are restrained inthe supine position with the penile sheath retracted until penileresponses are observed. Some animals require additional days ofconditioning before they are completely acclimated to the procedures;non-responders are removed from further evaluation. After any handlingor evaluation, animals are given a treat to ensure positivereinforcement.

2) Ex Copula Reflex Tests. Rats are gently restrained in a supineposition with their anterior torso placed inside a cylinder of adequatesize to allow for normal head and paw grooming. For a 400–500 gram rat,the diameter of the cylinder is approximately 8 cm. The lower torso andhind limbs are restrained with a non-adhesive material (vetrap). Anadditional piece of vetrap with a hole in it, through which the glanspenis will be passed, is fastened over the animal to maintain thepreputial sheath in a retracted position. Penile responses will beobserved, typically termed ex copulu genital reflex tests. Typically, aseries of penile erections will occur spontaneously within a few minutesafter sheath retraction. The types of normal reflexogenic erectileresponses include elongation, engorgement, cup and flip. An elongationis classified as an extension of the penile body. Engorgement is adilation of the glans penis. A cup is defined as an intense erectionwhere the distal margin of the glans penis momentarily flares open toform a cup. A flip is a dorsiflexion of the penile body.

Baseline and/or vehicle evaluations are conducted to determine how andif an animal will respond. Some animals have a long duration until thefirst response while others are non-responders altogether. During thisbaseline evaluation, latency to first response time, number and type ofresponses are recorded. The testing time frame is 15 minutes after thefirst response.

After a minimum of 1 day between evaluations, these same animals areadministered a compound of formula I at 20 mg/kg and evaluated forpenile reflexes. All evaluations are videotaped and scored later. Dataare collected and analyzed using paired 2 tailed t-tests to comparedbaseline and/or vehicle evaluations to drug treated evaluations forindividual animals. Groups of a minimum of 4 animals are utilized toreduce variability.

Positive reference controls are included in each study to assure thevalidity of the study. Animals can be dosed by a number of routes ofadministration depending on the nature of the study to be performed. Theroutes of administration includes intravenous (IV), intraperitoneal(IP), subcutaneous (SC) and intracerebral ventricular (ICY).

E. Models of Female Sexual Dysfunction:

Rodent assays relevant to female sexual receptivity include thebehavioral model of lordosis and direct observations of copulatoryactivity. There is also a urethrogenital reflex model in anesthetizedspinally transected rats for measuring orgasm in both male and femalerats. These and other established animal models of female sexualdysfunction are described in McKenna, et al., Am. J. Physiol.,(Regulatory Integrative Comp. Physiol 30):R1276–R1285, 1991; McKenna, etal., Pharm. Bioch. Behav., 40:151–156, 1991; and Takahashi, et al.,Brain Res., 359:194–207, 1985.

Preparation of the Compounds of the Invention

Preparation of the compounds of the present invention may be carried outvia sequential or convergent synthetic routes. The skilled artisan willrecognize that, in general, the three domains of a compound of formula Iare connected via amide bonds. The B and C domains are optionallyconnected via a reduced or partially reduced amide bond (e.g., viareductive amination). The skilled artisan can, therefore, readilyenvision numerous routes and methods of connecting the three domains viastandard peptide coupling reaction conditions.

The phrase “standard peptide coupling reaction conditions” meanscoupling a carboxylic acid with an amine using an acid activating agentsuch as EDC, dicyclohexylcarbodiimide, and benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate in a inert solvent suchas DCM in the presence of a catalyst such as HOBT. The uses ofprotective groups for amine and carboxylic acids to facilitate thedesired reaction and minimize undesired reactions are well documented.Conditions required to remove protecting groups which may be present canbe found in Greene, et al., Protective Groups in Organic Synthesis, JohnWiley & Sons, Inc., New York, N.Y. 1991.

CBZ, Boc or FMOC protecting groups are used extensively in thesynthesis, and their removal conditions are well known to those skilledin the art. For example, removal of CBZ groups can he achieved bycatalytic hydrogenation with hydrogen in the presence of a noble metalor its oxide such as palladium on activated carbon in a protic solventsuch as ethanol. In cases where catalytic hydrogenation iscontraindicated by the presence of other potentially reactivefunctionality, removal of CBZ can also be achieved by treatment with asolution of hydrogen bromide in acetic acid, or by treatment with amixture of TFA and dimethylsulfide. Removal of Boc protecting groups iscarried out in a solvent such as methylene chloride, methanol or ethylacetate with a strong acid, such as TFA or HCl or hydrogen chloride gas.

The compounds of formula I, when exist as a diastereomeric mixture, maybe separated into diastereomeric pairs of enantiomers by fractionalcrystallization from a suitable solvent such as methanol, ethyl acetateor a mixture thereof. The pair of enantiomers thus obtained may beseparated into individual stereoisomers by conventional means by usingan optically active acid as a resolving agent. Alternatively, anyenantiomer of a compound of the formula I may be obtained bystereospecific synthesis using optically pure starting materials orreagents of known configuration.

The compounds of the present invention can be prepared according to theprocedure of the following schemes and examples, which may furtherillustrate details for the preparation of the compounds of the presentinvention. The compounds illustrated in the examples are, however, notto be construed as forming the only genus that is considered as thepresent invention.

In coupling procedure 1, an appropriate A domain (e.g., piperazine) iscoupled to B domain (e.g., D-Boc-p-Cl-Phe-OH) in the presence ofEDC/HOBt followed by Boc deprotection in the presence of TFA. Thecoupled AB compound is then coupled to an appropriate C domain followedby deprotection of Boc group and salt formation. Alternatively, when Cdomain is not protected with Boc group, the final compound can beobtained without the deprotection step.

In coupling procedure 2, an appropriate A domain (e.g., piperazine) iscoupled to an appropriate BC domain in the presence of HATU followed bydeprotection of Boc group in the presence of TFA and salt formation.Alternatively, when BC domain is not protected with Boc group, the finalcompound can be obtained without the deprotection step.

In coupling procedure 3, an appropriate A domain is coupled to anappropriate BC domain in the presence of EDC/HOBT followed bydeprotection of Boc group in the presence of TFA and salt formation.

In coupling procedure 4, an appropriate AB domain is coupled to anappropriate C domain in the presence of EDC/HOBT followed bydeprotection of Boc group in the presence of TFA and salt formation.Alternatively, when C domain is not protected with Boc group, the finalcompound can be obtained without the deprotection step.

In coupling procedure 5, an appropriate A domain is coupled to anappropriate BC domain (protected or non-protected BC domain) in thepresence of HATU followed by deprotection of Boc group in the presenceof TFA and salt formation

In coupling procedure 6, an appropriate A domain is coupled to anappropriate BC domain (protected or non-protected BC domain) in thepresence of HATU followed by deprotection of CF₃CO group in the presenceof NH₃/MeOH and salt formation. Coupling procedure 6 is preferred toprepared piperidine derivatives of the present invention.

Alternatively, EDC/HOAT, EDC/HOBT or DCC/HOBT can be used when A domainis coupled with B domain.

Generally, the starting material of Boc-protected piperazine (A domain)can be deprotected in the presence of TFA/CH₂Cl₂, HCl/EtOAc,HCl/dioxane, or HCl in MeOH/Et₂O with or without a cation scavenger,such as dimethyl sulfide (DMS) before being subjected to the couplingprocedure. It can be freebased before being subjected to the couplingprocedure or in some cases used as the salt.

A suitable solvent such as CH₂Cl₂, DMF, THF or a mixture of the abovesolvents can be used for the coupling procedure. Suitable base includestriethyl amine (TEA), diisopropyethyl amine (DIPEA), N-methymorpholine,collidine, or 2,6-lutidine. Base may not be needed when EDC/HOBt isused.

Generally after the reaction is completed, the reaction mixture can bediluted with an appropriate organic solvent, such as EtOAc, CH₂Cl₂, orEt₂O, which is then washed with aqueous solutions, such as water, HCl,NaHSO₄, bicarbonate, NaH₂PO₄, phosphate buffer (pH 7), brine or anycombination thereof. The reaction mixture can be concentrated and thenbe partitioned between an appropriate organic solvent and an aqueoussolution. The reaction mixture can be concentrated and subjected tochromatography without aqueous workup.

Protecting group such as Boc or CBZ, FMOC, CF₃CO and H₂/Pd—C can bedeprotected in the presence of TFA/CH₂Cl₂, HCl/EtOAc, HCl/dioxane, HClin MeOH/Et₂O, NH3/MeOH, TBAF or H₂/Pd—C with or without a cationscavenger, such as thioanisole, ethane thiol and dimethyl sulfide (DMS).The deprotected amines can be used as the resulting salt or arefreebased by dissolving in CH₂Cl₂ and washing with aqueous bicarbonateor aqueous NaOH. The deprotected amines can also be freebased by SCX ionexchange chromatography.

The compounds of the present invention can be prepared as salt, such asTFA, hydrochloride or succinate salts by using known standard methods.

In the Schemes, Preparations and Examples below, various reagent symbolsand abbreviations have the following meanings:

BINAP 2,2′-Bis(diphenylphosphino)-1,1′-binaphthyl Boc t-butoxycarbonylCBZ benzyloxycarbonyl DCM dichloromethane DEAD diethyl azodicarboxylateDIAD diisopropyl azodicarboxylate DIPEA diisopropylethylamine DMAP4-dimethylamino pyridine DMF N,N-dimethylformamide DMSOdimethylsulfoxide eq. equivalent(s) EDC1-(3-dimethylaminopropyl)-3-ethylcarbodiimide HCl ESI-MS electron sprayion-mass spectroscopy Et ethyl EtOAc ethyl acetate FMOC9-Flurorenylmethyl carbamate HATUO-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate HOAT 1-hydroxy-7-azabenzotriazole HOBT1-hydroxybenzotriazole hydrate HPLC high performance liquidchromatography HRMS high resolution mass spectroscopy h (hr) hour(s)LRMS low resolution mass spectroscopy Me methyl Ms methanesulfonyl NMM4-methyl morpholine Pd₂(dba)₃ tris(dibenzylideneacetone) dipalladium(0)Ph phenyl Phe phenylalanine Pr propyl r.t. room temperature TBAFtetrabutylammonium fluoride TBS tertbutyldimethylsilyl TFAtrifluoroacetic acid TEA triethylamine THF tetrahydrofuran Tic1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid TLC thin-layerchromatographyReaction Scheme for Preparation of “A Domain”

The A domains of the present invention, in general, may be prepared fromcommercially available starting materials via known chemicaltransformations. The preparation of A domain of the compound of thepresent invention is illustrated in the reaction scheme 2 below.

The present invention also provides a novel process for preparingcertain intermediates and/or compounds of the invention as shown inReaction Schemes 3–5.

As shown in Reaction Scheme 3, a convergent synthesis of a keyintermediate isoindoline (5) via a Heck coupling, followed by areductive amination, a ring cyclization and a resolution has beendeveloped. Also, alternate asymmetric approaches including asymmetricMichael addition and asymmetric hydrogenation have also been developedto prepare compounds of the invention and/or intermediates thereof.

As shown in Reaction Scheme 3, the isoindoline compounds of the presentinvention may be prepared from 2-halobenzaldehyde 1 or substitutedanalog thereof. Preferred starting material is 2-bromobenzaldehyde orsubstituted analog thereof. Pd-mediated Heck coupling of2-bromobenzaldehydes 1 with for example, methyl acrylate, providedalpha,beta-unsaturated methyl esters 2, which undergoes reductiveamination to give amines, 3 (or carbamates where R₁ is for example,Boc). Various Heck coupling reagents and conditions were found suitableto effect the coupling reaction. Suitable catalysts and ligands includePd(OAc)₂/PPh₃, Pd(OAc)PPh₃/BU₄NBr, Pd(PPH₃)₂Cl₂/CUI, Pd(OAC)₂/P(O-Tol)₃.Suitable solvent or solvent systems for the Heck coupling reactioninclude DMF, toluene and ethyl acetate. More preferred base istriethylamine.

Reductive amination of the aldehyde functionality of 2 to amines isaccomplished in good yields by reaction with benzylamine oralpha-methylbenzylamine in acidic conditions, followed by in situreduction of the incipient imines with NaCNBH₃ at about pH 5. Otherreducing agents including Na(OAc)₃BH and NaBH/H may also be used toeffect reduction of the incipient imines. Interestingly, the resultingamines immediately cyclized to the isoindoline compounds under the sameacidic conditions for the reduction. Direct preparation of compound 4may also be effected by use of BocNH₂ instead of benzylamine in thereductive amination step. Screening of various reducing agentsdemonstrated that the combination of Et₃SiH and TFA in CH₃CN representsthe preferred method for effecting reductive amination using BocNH₂.

The N-Boc isoindolinecarboxylic acid 5 may also be prepared from 3 asthe carbamate, by an intra-molecular Michael addition and esterhydrolysis. The resolution of the isoindolinecarboxylic acids 4 bycrystallization afforded enantio-pure compounds 5.

Two alternate asymmetric approaches have also been developed for thesynthesis of isoindolinecarboxylic acid 5 i.e., asymmetric Michaeladditions and asymmetric hydrogenation. In the asymmetric Michaeladdition approach, alpha-methylbenzyl amine is used as a chiralauxiliary to induce the enantio-selectivity. In the asymmetrichydrogenation approach, compound 4′ could be converted to 5stereoselectively in the presence of chiral ligands.

Finally the coupling of the isoindolines 5 with the “B” domain piece,i.e., D-Cl-Phe to afford compound 6 (“BC” piece) is accomplished bystandard amino acid coupling reactions such as, for example, by the useof EDC or EDCI or other activating agents in the presence of suitable isdimethylaminopyridine (DMAP). The product (6) is then coupled with an“A” domain piece as discussed herein to afford the target MC4R agonistcompound of formula I by coupling reactions known to one of skill in theart.

Preferably, the isoindole or other “C” domain piece is coupled to an“AB” coupled domain piece to form the compound of formula I.

As shown in Reaction Scheme 4, m-tyrosine ester or analogs, includingsubstituted analogs thereof, may be esterified by forming the acidhalide followed by nucleophilic displacement of halide by the alkoxygroup from an alcohol, i.e., methanol or ethanol. Where thionyl chlorideor other halide source is used the product may be isolated as the acidaddition salt (2). The resulting ester (2) is subjected to aPictet-Spengler reaction by heating with a suitable ketone or aldehydein refluxing conditions. For example, an unsubstituted isoquinolinebackbone (3) may be formed by employing formaldehyde in thepictet-Spengler reaction. On the other hand, a gem-dimethyl substitutedisoquinoline wherein R¹¹ is methyl, may be formed by using acetone asthe ketone source and solvent. Other less reactive substituents may besubstituted as the R¹¹ group for the practice of the present invention.

The product isoquinoline (3) may be isolated preferably as the acidaddition salt. Where m-tyrosine is used as the starting material, thefree hydroxyl group is removed first by protection/activation with agood leaving group such as, for example, reaction with triflic anhydride(trifluoromethane sulfonic anhydride) or methanesulfonic acid to formthe triflate or mesylate in the presence of a base. The triflate is apreferred group used to set up the compound (3) for deoxygenationbecause of the extra electron withdrawing effect of the trifluoromethanesubstituent. The deoxygenation reaction is effected by hydrogenation atpressures of about 50 psi. The product (4) may be isolated as the acidaddition salt. The product (4) is hydrolyzed under basic conditions toafford the acid salt. Suitable bases for the above hydrolysis includeaqueous sodium hydroxide, potassium hydroxide and sodium lithiumhydroxide. The reaction is preferably performed in a mixture of aqueousand organic solvents. An exotherm during addition of base may beregulated (i.e., less than about 35° C.) to avoid overheating or“runaway reactions.” The reaction product may be isolated by aqueouswork up. Alternatively, the entire mixture may be concentrated andwashed with organic solvents to afford the desired product (6) aftercrystallization.

The product (6) is then reacted with a “B” domain substrate such as, forexample, 4-chloro-D-phenylalanine as described previously and in theexperimental section. The resulting “BC” combination product is thenreacted with an “A” domain piece to form the respective compound offormula I. Alternatively, the product (6) may be reacted with an “AB”domain combination product to afford a compound of formula I.

One of skill is aware that certain protections and deprotections ofintermediates in Reaction Scheme 4, to form the carbamate, substitutedamine or free amine at the isoquinolinyl nitrogen are possible andcontemplated as within the scope of this invention. Unless otherwisespecified, reagents and procedures for effecting the reactions describedherein are known to one of skill in the art and may be found in generalreference texts such as Advanced Organic Chemistry by J. March, 5^(th)edition, Wiley Interscience Publishers, New York, N.Y., and referencestherein.

In an alternate procedure, the isoquinoline product i.e., compound (3)or (5) including their N-protected analogs may be resolved by reactionwith a resolving agent such as for example, L-tartaric acid,dehydroabietylamine or other resolving agents known to one of skill inthe art.

Alternatively, asymmetric analogs of product (6) may be prepared byusing asymmetric starting materials. For example, L-DOPA may be used inplace of m-tyrosine ester in reactions essentially similar to thosedescribed and illustrated in Reaction Scheme 4 and in the examples, toafford the asymmetric analog of compound (6).

Tetrahydroisoquinoline acetic acid derivatives may be prepared andutilized as shown in Reaction Scheme 5 below:

As shown in Reaction Scheme 5, a compound of formula 10a wherein X ishalogen, preferably bromo or chloro, and R and R¹¹ are as definedpreviously, and which is obtained commercially or prepared fromcommercial starting materials is reacted with cyanomethylethylacetate toafford a compound of formula 10b. The compound of formula 10b may beprotected as the compound 10c with a suitable protecting group (Pg) andthen subjected to hydrogenation conditions including for exampleasymmetric hydrogenation to form a compound of formula 10d, which may bechiral (depending on hydrogenation conditions, i.e., asymmetric versusnon-asymmetric hydrogenation). The compound of formula 10d orstereoisomer thereof, is reacted with a B-domain piece such as, forexample, 4-chloro-D-phe to afford a BC piece (10e). The compound offormula 10e is then reacted with an A-domain piece to afford a compoundof formula I. The details of the specific reaction steps are similar toor analogous to reactions taught herein, and in the experimentalsection. Furthermore, one of skill in the art is aware of that suchintermediate reactions as hydrolysis and deprotection may be necessaryto achieve optimum yields in certain steps of the scheme as shown. Oneof skill in the art is also aware of further common manipulations suchas N-alkylation, or N-acylation, and alkylations on the benzene ring toafford other compounds of formula I.

The following describes the detailed examples of A Domain preparation.

Preparation of 1ADimethyl-[1-methyl-2-(2-piperazin-1-yl-phenoxy)-R-ethyl]-amine

Step 1: [2-(2-Bromo-phenoxy-1-R-methyl-ethyl]-carbamic acid tert-butylester

The o-bromophenol (1.98 g, 11.4 mmol) andR-2-hydroxy-1-methyl-ethyl-carbamic acid tert-butyl ester (2 g, 9.92mmol) were placed in THF (60 mL) with triphenylphosphine (4.5 g, 17.1mmol), and the system was cooled to about 0° C. DIAD (3.37 mL, 17.1mmol) was added portion-wise over 30 minutes. The mixture was warmed tor.t. and stirred for about 16 hours. The mixture was diluted with ether(200 mL) and water was added (100 mL). The mixture was washed with 5NNaOH (100 mL) and extracted with ether and concentrated. A solutionethyl acetate/hexanes was added, and the triphenylphosphine oxide wascrystallized and filtered away. Chromatography on silica gel (ethylacetate/hexanes) afforded about 2.78 g of the product (74%) as clearoil. MS found: 230.0 (M-Boc)

Step 2: 2-(2-bromo-phenoxy)-1-R-methyl-ethylamine

The compound of Step 1 (2.78 g, 8.42 mmol) was placed in DCM (10 mL) andTFA (10 mL) was added. The mixture was stirred at r.t. for about 16hours and then concentrated and subjected to SCX anion exchangechromatography for purification. About 1.84 g of the title compound wasobtained as clear oil (95%). MS found 230.0

Step 3: [2-(2-Bromo-phenoxy)-1-R-methyl-ethyl]-dimethyl amine

The compound of Step 2 (1.84 g, 7.99 mmol), formaldehyde (1.7 mL of 37%aq.), sodium triacetoxy borohydride (9.5 g, 44.7 mmol), and acetic acid(5.5 mL) were mixed together and stirred in dichloroethane (36 mL) atr.t. for about 4 hours. The mixture was diluted with DCM, quenched with1N NaOH (100 mL), and then the layers separated. The aqueous layer wasback extracted with ether (50 mL) and the organic phases were combined,which was then dried, filtered and concentrated. Chromatography onsilica gel afforded about 714 mg of the product (35%).

MS found 258.0

Step 4: The compound of Step 3 (714 mg, 2.72 mmol), piperazine (283 mg,3.26 mmol), Pd₂(dba)₃ (126 mg, 0.14 mmol), BINAP (255 mg, 0.41 mmol),and sodium tert-butoxide (368 mg, 3.81 mmol) were mixed together,degassed and toluene(20 mL) was added. The mixture was heated to 100° C.for about 16 hours. The mixture was cooled to r.t., diluted with ether(50 mL), filtered through celite and concentrated. The residue wassubjected to SCX purification, and the resultant oil was chromatographedon silica gel (MeOH (NH₃2M)/DCM) to give the final compound (243 mg,34%) as yellow oil. MS found 264.2, M+1

Preparation of 2ADimethyl-[1-methyl-2-(2-piperazin-1-yl-phenoxy)-S-ethyl]-amine

The title compound was prepared in a manner similar to Preparation 1Aexcept that S-2-hydroxy-1-methyl-ethyl-carbamic acid tert-butyl esterwas used.

MS found 264.2, M+1

Preparation of 3A4-[1-Methyl-2-(2-piperazin-1-yl-phenoxy)-R-ethyl]-morpholine

Step 1: 4-[2-(2-Bromo-phenoxy-1-R-methyl-ethyl]morpholine

2-(2-Bromo-phenoxy)-1-R-methyl-ethylamine (Preparation 1, Step 2)(1 g,4.35 mmol), 1-chloro-2-(2-chloro-ethoxy)-ethane (809 mg, 5.66 mmol),potassium iodide (145 mg, 0.87 mmol), potassium carbonate (2.4 g, 17.4mmol) and ethanol (40 mL) were mixed together and heated at reflux forabout 4 days. The mixture was concentrated to an oil, and the oil wastaken up in DCM and washed with water. The organic fraction was driedand concentrated. Chromatography on silica gel (MeOH/DCM) yielded theproduct (792 mg, 61%) as clear oil. MS found: 300.0 (M+1)

Step 2: The title compound was prepared by using Buchwald condition frompiperazine and 4-[2-(2-Bromo-phenoxy-1-R-methyl-ethyl]morpholine. MSfound 306.2, M+1

Preparation of 4A(R)-Ethyl-[1-methyl-2-(2-piperazin-1-yl-phenoxy)-ethyl]-amine

Step 1: (R)-[2-(2-Bromo-phenoxy)-1-methyl-ethyl]-ethyl-amine

(R)-2-(2-bromophenoxy)-1-methylethylamine, (Preparation 1A, Step 2)(1.68 g, 7.3 mmol) was dissolved in DMF (16 ml). The mixture was stirredwith bromoethane (3.18 ml, 43 mmol) and K₂CO₃ (7.60 g, 55 mmoles) atr.t. under N₂ for about 3 days. The mixture was concentrated, dissolvedin EtOAc, washed with water and brine, dried over Na₂SO₄, and filtered.Solvent was removed and the residue was purified by flash chromatography(silica gel, 5% 2M NH₃ in MeOH/CH₂Cl₂) to give about 2.08 g of a mixtureof diethyl and monoethyl amine compounds with the mono ethyl as theminor product.

MS ES MH+286/288 (1:1) (di) and MH+258/260 (1:1) (mono)

Step 2: (R)-Ethyl-[1-methyl-2-(2-piperazin-1-yl-phenoxy)-ethyl]-amine

The compounds of Step 1 (0.88 mg, 3.1 mmoles) was dissolved in drytoluene (8 ml) and then piperazine (0.34 g, 4.0 mmoles), Pd₂(dba)₃(0.140 g, 0.15 mmol), BINAP (0.29 g, 0.46 mmol) and sodium tert-butoxide(0.41 g, 4.3 mmol) were added. The mixture was degassed and heated toabout 85° C. for about 16 hours. The mixture was allowed to cool tor.t., and diluted with THF. The mixture was filtered through celite andthen concentrated. The residue was purified by flash chromatography(silica gel, 5% 2N NH₃ in MeOH/CH₂Cl₂) to give a mixture (0.34 g) ofmono- and di-ethyl compound, mono ethyl being as the minor product.

MS ES MH+ 292 (1:1) (di) and MH+ 264 (1:1) (mono)

Preparation of 5AN-Ethyl-N-[1-methyl-2-(2-piperazin-1-yl-phenoxy)-R-ethyl]-Methanesulfonamide

Step 1: N-[2-(2-Bromo-phenoxy)-1-R-methyl-ethyl]-acetamide

2-(2-bromo-phenoxy)-1-R-methyl-ethylamine (Preparation 1A, Step 2) (1.44g, 6.26 mmoles) was mixed with acetic anhydride (10 ml) and stirred atr.t. for an hour and then stirred another hour at 40–45° C. The mixturewas concentrated and water was added. The mixture was extracted withEtOAc, washed with NaHCO₃ and brine, and then dried over Na₂SO₄ andfiltered. Removal of solvent gave a residue, which was purified by flashchromatography (silica gel, EtOAc) to give the acetamide, (1.55 g, 91%).

MS ES MH+ 272/274 (1:1)

Step 2: [2-(2-Bromo-phenoxy)-1-R-methyl-ethyl]-ethyl-amine

N-[2-(2-Bromo-phenoxy)-1-R-methyl-ethyl]-acetamide (0.50 g, 1.8 mmol)was dissolved in THF (10 ml). BH₃-THP (1.5 M in THF, 3.6 ml, 5.4 mmol)was added and the mixture was heated at 60° C. for about an hour. Themixture was cooled to r.t., and then DIEA (1.2 ml) in MeOH (2.4 ml) wasadded followed by the addition of I₂ (0.91 g) in THF (5 ml). The mixturewas stirred for about 30 minutes and diluted with EtOAc. It was washedwith 1N Na₂S₂O₃, and brine, and then dried over Na₂SO₄ and filtered.Removal of solvent gave the crude ethylamine(0.54 g).

MS ES MH+ 258/260(1:1)

Step 3:N-[2-(2-Bromo-phenoxy)-1-R-methyl-ethyl]-N-ethyl-methanesulfonamide

[2-(2-Bromo-phenoxy)-1-R-methyl-ethyl]-ethyl-amine (0.54 g, 1.8 mmol)was dissolved in CH₂Cl₂ (10 ml). DIEA (0.63 ml, 3.6 mmol) and MsCl (0.17ml, 2.2 mmol) were added and the mixture was stirred at r.t. overnight.Additional MsCl (0.51 ml, 6.6 mmol) and DIEA (1.26 ml, 7.2 mmol) wereadded and the mixture was stirred at r.t. overnight. Additional MsCl(0.34 ml, 4.4 mmol) and DIEA (0.63 ml, 3.6 mmol) were added and thenstirred for another 9 hours. The mixture was diluted with EtOAc, washedwith diluted NaHCO₃ and brine, and then dried over Na₂SO₄ and filtered.Removal of solvent gave a residue, which was purified by flashchromatography (silica gel, EtOAc/hexane, ⅓) to give the sulfonamide(0.39 g, 64%, in two steps).

MS ES MH+ 336/338 (1:1)

Step 4: The final compound was prepared from piperazine andN-[2-(2-Bromo-phenoxy)-1-R-methyl-ethyl]-N-ethyl-methanesulfonamideusing Buchwald condition. MS ES MH+ 342

Preparation of 6ADimethyl-[2-(2-piperazin-1-yl-phenoxy)-R,S-propyl]-amine

The title compound was prepared in a manner similar to Preparation 1Aexcept that o-bromophenol was coupled with racemic1-dimethylamino-propan-2-ol.

MS found 264.2, M=+1

Preparation of 7A3-(2-Piperazin-1-yl-phenoxy)-S-pyrrolidine-1-carboxylic acid tert-butylester:

The title compound was prepared in a manner similar to Preparation 1Aexcept that o-bromophenol was coupled withR-3-hydroxy-pyrrolidine-1-carboxylic acid tert-butyl ester. MS found348.2, M+1

Preparation of 8A3-(2-Piperazin-1-yl-phenoxy)-R-pyrrolidine-1-carboxylic acid tert-butylester

The title compound was prepared in a manner similar to Preparation 7Aexcept that o-bromophenol was coupled withS-3-Hydroxy-pyrrolidine-1-carboxylic acid tert-butyl ester. MS found348.1 M+1

Preparation of 9A2-(2-Piperazin-1-yl-phenoxymethyl)-S-pyrrolidine-1-carboxylic acidtert-butyl ester

The title compound was prepared in a manner similar to Preparation 1Aexcept that o-bromophenol was coupled with Boc-L-prolinol. MS found362.3, M+1

Preparation 10A2-(2-Piperazin-1-yl-phenoxymethyl)-R-pyrrolidine-1-carboxylic acidtert-butyl ester

The title compound was prepared in a manner similar to Preparation 1Aexcept that o-bromophenol was coupled with Boc-D-prolinol. MS found362.2, M+1

Preparation of 11A4-[2-(1-tert-butoxycarbonyl-S-pyrrolidin-3-yloxy)-5-methyl-phenyl]piperazine

The title compound was prepared in a manner similar to Preparation 1Aexcept that 2-bromo-4-methyl phenol was coupled withN-boc-3-(R)-hydroxy-pyrrolidine.

LRMS (electrospray): 362.3 (M+1)

Preparation of 12A1-[5-Isopropyl-2-S-(pyrrolidin-3-yloxy)-phenyl]-piperazine

The title compound was prepared in a manner similar to Preparation 11Aexcept that 2-Bromo-4-isopropyl-phenol was coupled withN-boc-3-(R)-hydroxy-pyrrolidine.

Preparation of 13A1-[2-(1-Methyl-S-piperidin-3-yloxy)-phenyl]-piperazine

Step 1: R-piperidin-3-ol HCl salt (15 g, 109 mmol) was placed inDCM/water (500 mL of 1/1 mixture) with potassium carbonate (30.1 g, 218mmol), and di-t-butyl dicarbonate (26.2 g, 120 mmol) was added with offgassing. The mixture was stirred at r.t. overnight and then diluted withDCM (400 mL) and washed with water (2×200 mL). The organic fraction wasdried and concentrated to give white crystalline solids (23.2 g).

Step 2: The compound obtained in Step 1 (10 g, 49.68 mmol), orthobromophenol (7.14 g, 41.4 mmol) and triphenylphosphine (19.55 g, 74.5mmol) were placed in THF (100 mL), and the mixture was cooled to 0° C.DIAD (14.7 mL, 74.5 mmol) was added dropwise for about 30 minutes, andthe mixture was warmed to r.t. and stirred for about 16 hours. Themixture was diluted with ether (500 mL) and water was added (50 mL).Then the organic portion was washed with 5N NaOH (500 mL), and theaqueous phase was extracted with ether (500 mL) and concentrated. Theconcentrated oil was taken up in EtOAc/hexanes and thetriphenylphosphine oxide crystallized. The slurry was filtered, and thefiltrate was concentrated and chromatograped on silica gel to give theproduct (6.65 g, 45%) as a clear oil.

Step 3: The compound obtained in Step 2 (1 g, 2.81 mmol) was placed inDCM/TFA (20 mL of 1/1 mixture), and the mixture was stirred at r.t. forabout 16 hours. The mixture was concentrated and subjected to SCX anionexchange chromatography to give the product (775 mg) as a clear oil. MSfound 256.

Step 4: The compound of Step 3 (8.63 g, 33.7 mmol), formaldehyde (7.06mL of 37% aq.), sodium triacetoxy borohydride (40 g, 188.72 mmol) andglacial acetic acid (23.1 mL, 404.4 mmol) were mixed in dichloroethane(153 mL), and the mixture was stirred at r.t. for about 24 hours. Themixture was diluted with DCM (200 mL) and quenched with IN NaOH (200mL), and then the layers were separated. The aqueous layer was backextracted with ether (200 mL), and the organic phases were combined,dried, filtered and concentrated. Chromatography gave the product (8.18g, 90%) as a clear oil. MS found 270.02

Step 5: The compound of Step 4 (4.28 g, 15.84 mmol), piperazine (1.64 g,19.01 mmol), Pd₂(dba)₃ (725 mg, 0.792 mmol), BINAP (1.48 g, 2.38 mmol)and sodium tert-butoxide (2.13 g, 22.22 mmol) were mixed together andtoluene (100 mL) was added. The mixture was heated to 100° C. for about5 hours. The mixture was cooled to r.t., diluted with ether (100 mL),filtered through celite and concentrated. The residue was subjected toSCX purification, and the resultant oil was chromatographed to give thefinal compound (2.12 g, 49%) as a yellow oil. MS found 276.2

Preparation of 14A1-{2-[1-(2,2,2-Trifluoro-ethyl)-S-piperidin-3-yloxy]-phenyl}-piperazine

Step 1:1-[3-(2-Bromo-phenoxy)-S-piperidin-1-yl]-2,2,2-trifluoro-ethanone

About 1.0 g (3.9 mmol) of 3-(2-Bromo-phenoxy)-piperidine was dissolvedin 20 ml THF under nitrogen and 1.7 ml (12.0 mmol) trifluoraceticanhydride was added dropwise. The mixture was stirred for about 30minutes and concentrated in vacuo followed by the addition of ethylacetate. The mixture was washed with water and brine, and then driedwith sodium sulfate and concentrated to dryness. The resulting oil waschromatographed on silica by eluting with 10% ethyl ethyl acetate/90%hexane to afford about 1.25 g of the title compound. Mass MH⁺ 353

Step 2: 3-(2-Bromo-phenoxy)-1-(2,2,2-trifluoro-ethyl)-S-piperidine

About 1.25 g (2.13 mmol) of compound from Step 1 was dissolved in 14 mlTHF under nitrogen and 0.45 ml (2.13 mmol) borontrifluoride etherate wasadded. The mixture was heated to reflux and 5.3 ml of (6.39 mmol) boranedimethylsulfide was added dropwise over 10 minutes. The mixture wasstirred for about 45 minutes and the sulfide was allowed to boil off.After cooling, about 6–7 ml 6N HCl was added and the flask was heated toabout 40° C. The mixture was made basic with 2N NaOH and then ethylacetate was added. The organic layer was separated, washed with brine,dried with sodium sulfate and concentrated to dryness. The resulting oilwas chromatographed on silica by eluting with 20% ethyl ethylacetate/80% hexane to afford about 1.13 g of the title compound. MassMH⁺ 339

Step 3: The final compound was prepared by coupling the compound of Step2 with piperazine under Buchwald condition.

Preparation of 15A 3-(2-Piperazin-phenoxy)-R-piperidine-1-carboxylicacid tert-butyl ester

Sodium hydride (2.99 g, 74.55 mmol) (60% in mineral oil) was slurred inDMF (20 mL) and heated to 65° C. To the slurry was added 1 (5 g, 24.85mmol) in DMF (25 mL) dropwise over 30 minutes. The mixture was stirredat 65° C. for about an hour. Bromofluorobenzene (5.5 mL, 49.7 mmol) inDMF (5 mL) was added dropwise, and the mixture was stirred at 65° C. forabout 16 hours. The reaction was diluted with water and concentrated toan oily solid, which was then extracted between water (200 mL) and 1/1EtOAc/hexanes (200 mL). The organic layer was dried, filtered andconcentrated. Chromatography gave the compound 2 (6.35 g, 72%) as aclear oil.

Step 2: Compound 2 (2 g, 5.61 mmol), piperazine (581 mg, 6.74 mmol),Pd₂(dba)₃ (256 mg, 0.28 mmol), BINAP (523 mg, 0.84 mmol), and sodiumtert-butoxide (755 mg, 7.85 mmol) were mixed together and toluene (50mL) was added. The mixture was heated to 100° C. for about 5 hours. Themixture was allowed to cool to r.t, which was then diluted with ether(100 mL), filtered through celite and concentrated. The residue wassubjected to SCX purification, and the resultant oil was chromatographedto give the final compound (1.56 g, 77%) as a yellow oil. MS found362.2, M+1

Preparation of 16A1-[2-(1-methyl-R-piperidin-3-yloxy)-phenyl)-phenyl]-piperazine

The title compound was prepared in a manner similar to Preparation 13Aexcept that R-3-(2-Bromo-phenoxy)-piperidine was used.

Preparation of 17A Synthesis of4-[2-(N-boc-piperidin-4-yloxy)-phenyl]-piperazine

Step 1: N-boc-4(2-bromo-phenoxy)-piperidine

To a slurry of NaH (60% dispersion in oil, 5.9 g, 148.5 mmol, 3.0 eq.)in DMP (40 mL) at 65° C. under N₂ was added a solution ofN-boc-4-hydroxy piperidine (10 g, 49.5 mmol, 1.0 eq.) in DMF (50 mL).After stirring for about 2 hours, a solution of 1-bromo-2-fluoro benzene(11.0 mL, 99.0 mmol, 2.0 eq.) in DMF (10 mL) was added dropwise. Theresulting mixture was stirred at 65° C. overnight. The mixture wasdiluted with EtOAc (250 mL) and washed with water (100 mL) and brine(100 mL). The aqueous layer was extracted with EtOAc (3×). The combinedorganic extracts were dried (Na₂SO₄), filtered and concentrated.Purification by flash chromatography (250 g SiO₂, linear gradient, 40mL/min, 0% to 30% EtOAc/Hexane over 20 minutes and 30% EtOAc for 13minutes) afforded title compound (14.2 g, 39.8 mmol, 81%). LRMS(electrospray): 358.0.

Step 2: To a solution of N-boc-4-(2-bromo-phenoxy)-piperidine (13.4 g,39.6 mmol, 1.0 eq.), piperazine (9.73 g, 113 mmol, 3.0 eq.), Pd₂dba₃(1.72 g, 1.88 mmol, 0.05 eq.), BINAP (3.5 g, 5.65 mmol, 0.15 eq.) intoluene (150 mL) was added NaOtBu (5.1 g, 52.6 mmol, 1.4 eq.). Themixture was heated to 95° C. and stirred at that temperature overnight.The slurry was diluted with EtOAc and filtered through a pad of celite.The filtrate was concentrated and purified by ion exchange (SCX, 10 g)column. Further purification by flash chromatography (250 g SiO₂, 40ml/min, linear gradient 0–10% 2.0 M NH₃ in MeOH/CH₂Cl₂ for 20 minutesand 10% 2.0 M NH₃ in MeOH/CH₂Cl₂ for 73 minutes) afforded the finalcompound (12.65 g, 93%). LRMS (electrospray): 362.7 (M+1).

Preparation of 18A3-(2-Piperazin-1-yl-phenoxy)-S-piperidine-1-carboxylic acid tert-butylester

Step 1: 3-(2-Bromo-phenoxy)-S-piperidine-1-carboxlic acid tert-butylester

The o-bromophenol (1.72 g, 9.92 mmol) and Boc-R-3-hydroxypiperidine (2g, 9.92 mmol) were placed in THF (60 mL) with triphenylphosphine (3.9 g,14.9 mmol), and the system was cooled to 0° C. DIAD (2.94 mL, 14.9 mmol)was added portion wise over 30 minutes and the mixture was warmed tor.t. and stirred for about 16 hours. The mixture was diluted with ether(200 mL) and water was added (100 mL). The mixture was washed with 5NNaOH (100 mL), extracted with ether and concentrated. Ethylacetate/hexanes was added and the triphenylphosphine oxide wascrystallized and filtered away. The residue was chromatographed onsilica gel (ethyl acetate/hexanes) afforded the product (588 mg, 17%) asa clear oil. MS found: 256.0 (M-Boc)

Step 2: The compound of Step 1 (588 mg, 1.65 mmol), piperazine (171 mg,1.98 mmol), Pd₂(DBA)₃ (76 mg, 0.083 mmol), BINAP (154 mg, 0.248 mmol),and sodium tert-butoxide (222 mg, 2.31 mmol) were mixed together anddegassed followed by the addition of toluene (15 mL). The mixture washeated to 100° C. for about 16 hours. The mixture was cooled to r.t.,and diluted with ether (50 mL), filtered through celite andconcentrated. The residue was subjected to SCX purification, and theresultant oil was chromatographed on silica gel (10% MeOH (NH₃ 2M)/DCM)to afford the final compound (103 mg, 17%) as a yellow oil.

MS found 362.2, M=+1

Preparation of 19A 3-(2-Piperazin-1-yl-phenoxy)-piperidine-1-carboxylicacid tert-buyl ester

Step 1: 3-(2-Bromo-phenoxy)-piperidine-1-carboxylic acid tert-butylester

Ortho-bromophenol (1.72 g, 9.92 mmol) and racemicBoc-3-hydroxypiperidine (2 g, 9.92 mmol) were placed in THF (60 mL) withtriphenylphosphine (3.9 g, 14.9 mmol), and the mixture was cooled to 0°C. DIAD (2.94 mL, 14.9 mmol) was added portion wise over 30 minutes, andthe mixture was warmed to r.t. and stirred for about 16 hours. Themixture was diluted with ether (200 mL) and water was added (100 mL).The mixture was washed with 5N NaOH (100 mL), extracted with ether andconcentrated. Ethyl acetate/hexanes was added, and thetriphenylphosphine oxide was crystallized and filtered away. The residuewas chromatographed on silica gel (ethyl acetate/hexanes) to afford theproduct (1.2 g, 34%) as a clear oil. MS found: 256.0 (M-Boc)

Step 2: The compound of Step 1 (600 mg, 1.68 mmol), piperazine (174 mg,2.02 mmol), Pd₂(dba)₃ (77 mg, 0.084 mmol), BINAP (157 mg, 0.252 mmol),and sodium tert-butoxide (226 mg, 2.35 mmol) were mixed together anddegassed followed by the addition of toluene (20 mL). The mixture washeated to 100° C. for about 16 hours. The mixture was cooled to r.t, andthen diluted with ether (50 mL), filtered through celite andconcentrated. The residue was subjected to SCX purification, and theresultant oil was chromatographed on silica gel (MeOH (NH₃ 2M)/DCM) togive the final compound (311. mg, 51%) as a yellow foam. MS found 362.3,M+1

Preparation of 20A1-[3-(2-piperazin-1-yl-phenoxy)-R-piperdin-1-yl]-ethanone

Step 1:2,2,2-trifluoro-1-{4-[2-(R-piperidin-3-yloxy)-phenyl]-piperazin-1-yl}-ethanone

3-(2-piperazin-1-yl-phenoxy)-R-piperidin-1-carboxylic acid tert-butylester (1.56 g, 4.32 mmol) was dissolved in DCM (20 mL). Pyridine (1.37g, 17.28 mmol) and trifluoroacetic anhydride (1.81 g, 8.63 mmol) wereadded to the mixture and then stirred at r.t. for about 12 h. Themixture was subjected to an aqueous work up, and then dissolved inDCM/TFA (1/1, 20 mL), which was stirred at r.t. for about 4 hours. Thematerial was concentrated and chromatographed to afford the finalcompound (1.34 g, 86%) as a yellow foam. MS found: 358.1 M+1

Step 2: The compound of Step 1 was dissolved in acetic anhydride (10 mL)and triethyl amine (3 mL) was added. The mixture was stirred at r.t. forabout 6 hours and concentrated to dryness, which was then taken up inDCM (50 mL). The mixture was washed with 1N NaOH (50 mL), and theorganic fraction was dried and concentrated. The material was placed in7N NH3/MeOH (50 mL) and aged for about 48 hours. The resulting materialwas subjected to SCX purification and purified on silica gel to give thefinal compound (613 mg, 92%) as a pale yellow oil. MS found 304.2, M+1

Preparation of 21A1-[3-(2-piperazin-1-yl-phenoxy)-S-piperdin-1-yl]-ethanone

The title compound was prepared in a manner similar to Preparation 20Aexcept that 3-(2-piperazin-1-yl-phenoxy)-S-piperidin-1-carboxylic acidtert-butyl ester was used.

Preparation of 22A1-[3-(2-Piperazin-1-yl-phenoxy)-piperidin-1-yl]-propan-1-one

To2,2,2-trifluoro-1-{4-[2-(S-piperidin-3-yloxy)-phenyl]-piperazin-1-yl}-ethanone(prepared by following substantially similar procedure as described inPreparation 20A) (0.198 g, 0.552 mmol), TEA (0.085 ml, 0.608 mmol) inDCM (2 mL) at 0° C. and propionyl chloride (0.053 ml, 0.608 mmol) wasadded. The mixture was stirred at r.t. for about 2 hours. The mixturewas concentrated and DCM was added. The resulting solution was washedwith 1N NaOH, separated, dried over sodium sulfate, and concentrated togive yellow oily material. MS M+1 414.3.

The resulting amide was mixed with 7N NH₃ (10 ml) and aged at r.t. forthree (3) days. The solution was concentrated and purified through SCXcolumn and chromatographed on silica to afford about 94 mg of the finalcompound (54%). MS M+1 318.2.

Preparation of 23A2-Methyl-1-[3-(2-piperazin-1-yl-phenoxy)-piperidin-1-yl]-propan-1-one

The title compound was prepared in a manner similar to Preparation 22A.MS 332.2, M+H

Preparation of 24A 3-(2-Piperazin-1-yl-phenoxy)-piperidine-1-carboxylicacid methyl ester

The title compound was prepared in a manner similar to Preparation 22Aexcept that methyl chloroformate was used. MS 320.2, M+H

Preparation of 25A 3-(2-Piperazin-1-yl-phenoxy)-piperidine-1-carboxylicacid ethyl ester

The title compound was prepared in a manner similar to Preparation 22Aexcept that ethyl chloroformate was used. MS 334.2, M+H

Preparation of 26A 3-(2-Piperazin-1-yl-phenoxy)-piperidine-1-carboxylicacid isopropyl ester

The title compound was prepared in a manner similar to Preparation 22Aexcept that isopropyl chloroformate was used.

MS 348.2, M+H

Preparation of 27A1-[2-(1-Methanesulfonyl-piperidin-3-yloxy)-phenyl]-piperazine

The title compound was prepared in a manner similar to Preparation 22Aexcept that methanesulfonyl chloride was used.

MS 340.2, M+H

Preparation of 28A1-[2-(1-Ethanesulfonyl-piperidin-3-yloxy)-phenyl]-piperazine

The title compound was prepared in a manner similar to Preparation 22Aexcept that ethanesulfonyl chloride was used.

MS 354.2, M+H

Preparation of 29A1-{2-[1-(Propane-2-sulfonyl)-piperidin-3-yloxy]-phenyl}-piperazine

The title compound was prepared in a manner similar to Preparation 22Aexcept that isopropanesulfonyl chloride was used.

MS 368.2, M+H

Preparation of 30A3-(6-Fluoro-2-piperazin-1-yl-phenoxy)-piperidine-1-carboxylic acidtert-butyl ester

Step 1: 2-Bromo-3-fluoro-anisole

The 3-fluoro anisole (15 g, 119 mmol) was placed in THF (300 mL) andcooled to about −78° C., and then n-BuLi (89 mL 1.6 M in hexanes) wasadded while maintaining the temperature at around −70° C. The mixturewas stirred for about 15 minutes, and bromine (18.9 g, 119 mmol) wasadded over 10 minutes. The mixture was warmed to r.t. and stirredovernight. The mixture was quenched with water, diluted with ether (500mL) and partitioned between water/ether (1/1 300 mL). The water layerwas back extracted with ether (250 mL), and the combined organicfractions were dried, filtered and concentrated. The dark residue wasdistilled under house vacuum (product 115–130° C.) to recover about 13.8g of the product as a gray oil (57%).

Step 2: 2-bromo-3-fluorophenol

2-Bromo-3-fluoro anisole (10 g, 48.77 mmol) was dissolved in DCM (200mL) and cooled to about −78° C. To the solution was added BBr₃ (11.53mL, 121.9 mmol) in one portion. The mixture was warmed to r.t. andstirred overnight. The mixture was partitioned between water and DCM (50mL), and the aqueous layer was back extracted with DCM (200 mL). Thecombined organics were dried (NaSO₄), filtered and concentrated to giveabout 9.32 g of the product as a dark oil. MS found 188.9 (M−1)

Step 3: The final compound was prepared by using Mitsunobu followed byBuchwald condition from 2-bromo-3-fluorophenol and racemicN-boc-3-hydroxypiperidine.

MS found 380.3 M+1

Preparation of 31A2-Fluoro-5-(2-piperazin-1-yl-phenoxy)-piperidine-1-carboxylic acidtert-butyl ester

Step 1: 2-Chloro-6-fluoro phenol

2-Chloro-6-fluoro anisole (5 g, 31.13 mmol) was dissolved in DCM (300mL) and cooled to about −78° C. To the solution was added BBr₃ (7.35 mL,77.8 mmol) in one portion. The reaction was warmed to r.t, stirred forabout 16 hours and poured over ice to quench remaining BBr₃. Thematerial was partitioned between water and DCM (500 mL each). Theaqueous layer was back extracted with DCM (200 mL), and the combinedorganics were dried (NaSO₄), filtered and concentrated to give theproduct (4.60 g) as a dark oil.

MS found 145.0 M−1

Step 2: The title compound was prepared using Mitsunobu followed byBuchwald condition from 2-chloro-6-fluoro phenol and racemicN-boc-3-hydroxypiperidine.

MS found 380.3 M+1

Preparation of 32A 3-(2-Bromo-4-fluoro-phenoxy)-piperidine-1-carboxylicacid tert-butyl ester

The title compound was prepared by using Mitsonobu followed by Buchwaldcondition from 2-bromo-4-fluoro-phenol and racemicN-boc-3-hydroxypiperidine.

MS found: 380.3 M=+1

Preparation of 33A3-(5-Fluoro-2-piperazin-1-yl-phenoxy)-piperidine-1-carboxylic acidtert-butyl ester

The title compound was prepared by using Mitsunobu followed by Buchwaldcondition from 2-bromo-5-fluoro-phenol and racemicN-boc-3-hydroxypiperidine.

Preparation of 34A3-(2-Piperazin-1-yl-trifluoromethyl-phenoxy)-S-pyrrolidine-1-carboxylicacid tert-butyl ester

The title compound was prepared by using Mitsunobu followed by Buchwaldcondition from 2-bromo-4-trifluoromethyl phenol andN-boc-R-3-pyrrolidinol. MS 416.3 (M+1)

Preparation of 35A4-[2-(2-Morpholin-4-yl-ethoxy)-phenyl]-piperazine-1-carboxylic acidtert-butyl ester

Diisopropyl azodicarboxylate (0.48 mL, 2.4 mmol) was added dropwise to astirred solution of 4-(2-hydroxy-phenyl)-piperazine-1-carboxylic acidtert-butyl ester (0.56 g, 2 mmol), 2-morpholin-4-yl-ethanol (0.24 mL, 2mmol) and triphenylphosphine (0.63 g, 2.4 mmol) in THF (20 mL) cooled to0° C. The resulting solution was warmed to r.t. and stirred for about 16hours. The solvent was removed in vacuo and the residue purified bysilica gel chromatography (ethyl acetate) to afford the final compound(0.50 g, 64%) as a clear oil

¹H NMR (CDCl₃) δ 6.80–7.05 (m, 4H), 4.10–4.15 (m, 2H), 3.55–3.80 (m,8H), 3.00–3.10 (m, 4H), 2.75–2.85 (m, 2H), 2.60–2.70 (m, 4H), 1.55 (s,9H); TLC (SiO₂): R_(f)=0.25 (ethyl acetate)

Preparation of 36A4-[2-(1-Methyl-piperidin-4-yloxy)-phenyl]-piperazine-1-carboxylic acidtert-butyl ester

The title compound was prepared by using Mitsonobu condition from4-(2-hydroxyphenyl)-piperazine-1-carboxylic acid-tert-butyl ester andN-methylpiperidinol.

Preparation of 37A4-[2-(1-tert-Butoxycarbonyl-piperidin-4-yloxy)-phenyl]-piperazine-1-carboxylicacid benzyl ester

Diisopropyl azodicarboxylate (0.70 mL, 3.55 mmol) was added dropwise toa stirred solution of 4-(2-hydroxy-phenyl)-piperazine-1-carboxylic acidbenzyl ester (0.74 g, 2.4 mmol), 4-hydroxy-piperidine-1-carboxylic acidtert-butyl ester (0.48 g, 2.4 mmol) and triphenylphosphine (0.93 g, 3.6mmol) in THF (25 mL) cooled to 0° C. The resulting solution was warmedto r.t. and stirred for about 16 hours. The solvent was removed in vacuoand the residue purified by silica gel chromatography (20% ethyl acetatein hexanes) to afford the final compound (0.50 g, 43%) as a clear oil.

¹H NMR (CDCl₃) δ 6.80–7.35 (m, 9H), 5.25 (s, 2H), 4.45–4.55 (s, 1H),2.80–3.80 (m, 12H), 1.75–2.5 (m, 4H), 1.50 (s, 9H) TLC (SiO₂):R_(f)=0.30 (20% ethyl acetate in hexanes)

Preparation of 38A4-[2-(1-Methyl-piperidin-3-yloxy)-phenyl]-piperazine-1-carboxylic acidtert-butyl ester

The title compound was prepared in a manner similar to4-[2(2-Morpholin-4-yl-ethoxy)-phenyl]-piperazine-1-carboxylicacidtert-butyl ester except that4-(2-hydroxy-phenyl)-piperazine-1-carboxylic acid tert-butyl ester wascoupled to 1-methyl-piperidin-3-ol.

Preparation of 39A4-[2-(1-Methyl-piperidin-2-ylmethoxy)-phenyl]-piperazine-1-carboxylicacid tert-butyl ester

The title compound was prepared by using Mitsunobu condition from4-(2-hydroxy-phenyl)-piperazine-1-carboxylic acid tert-butyl ester and1-Methyl-2-piperidinemethanol.

Preparation of 40A4-[2-(1-Ethyl-pyrrolidin-3-yloxy)-phenyl]-piperazine-1-carboxylic acidtert-butyl ester

The title compound was prepared by using Mitsunobu condition from4-(2-hydroxy-phenyl)-piperazine-1-carboxylic acid tert-butyl ester and1-Ethyl-3-pyrrolidinol.

Preparation of 41A4-[2-(1-Methanesulfonyl-piperidin-3-yloxy)-phenyl]-piperazine-1-carboxylicacid tert-butyl ester

Step 1: 1-Methanesulfonyl-piperidin-3-ol

Piperidin-3-ol (5.0 g, 49.4 mmol) was dissolved in 1/1 THF/methylenechloride (100 mL). Triethylamine (17 mL, 123 mmol) was added, and thesolution was cooled to 0° C. Methanesulfonyl chloride (4.0 mL, 52 mmol)was added dropwise. The mixture was warmed to r.t. and stirredovernight. The reaction was diluted with ethyl acetate (800 mL), washedwith 1 N HCl (100 mL), water (75 mL), saturated aqueous sodiumbicarbonate (75 mL) and brine (75 mL), and then dried over anhydroussodium sulfate. The solvent was concentrated under reduced pressure andpurified via silica gel chromatography (ethyl acetate) to afford thetitle compound as a clear oil.

¹H NMR (CDCl₃) δ 3.83–3.96 (m, 1H), 3.40–3.53 (m, 1H), 3.23–3.37 (m,1H), 2.96–2.19 (m, 2H), 2.80 (s, 3H), 1.78–2.02 (m, 3H), 1.48–1.75 (m,1H).

Step 2: The title compound was prepared by using Mitsunobu conditionfrom 4-(2-Hydroxy-phenyl)-piperazine-1-carboxylic acid tert-butyl esterand 1-methanesulfonyl-piperidin-3-ol.

Preparation of 42A4-[2-(1-Acetyl-piperidin-4-yloxy)-phenyl]-piperazine-1-carboxylic acidtert-butyl ester

Step 1: 1-(4-Hydroxy-piperidin-1-yl)-ethanone

4-hydroxypiperidine (0.534 g, 5.3 mmol) was dissolved in methylenechloride (50 mL). The solution was cooled to −78° C. and triethylamine(1.10 mL, 7.95 mmol) was added. Acetyl chloride (0.33 mL, 4.77 mmol) wasadded dropwise to the mixture. The mixture was warmed to r.t.,concentrated under reduced pressure and purified using silica gelchromatography (50% ethyl acetate in hexanes) to give the title compound(0.540 g, 80%) as an oil.

¹H NMR (CDCl₃) δ 4.05–3.90 (m, 2H), 3.75–3.65 (m, 1H), 3.25–3.05 (m,2H), 2.25 (s, 1H), 2.05 (s, 3H), 1.90–1.75 (m, 2H), 1.60–1.40 (m, 2H).

Step 2: The title compound was prepared by using Mitsunobu conditionfrom 4-(2-hydroxy-phenyl)-piperazine-1-carboxylic acid tert-butyl esterand 1-(4-Hydroxy-piperidin-1-yl)-ethanone.

Preparation of 43A4-[2-(2-Pyrrolidin-1-yl-ethoxy)-phenyl]-piperazine-1-carboxylic acidtert-butyl ester

The title compound was prepared by using Mitsunobu condition from4-(2-hydroxy-phenyl)-piperazine-1-carboxylic acid tert-butyl ester and2-hydroxyethyl pyrrolidine.

Preparation 44A4-[2-(1-Methyl-pyrrolidin-3-yloxy)-phenyl]-piperazine-1-carboxylic acidtert-butyl ester

The title compound was prepared using Mitsunobu condition from4-(2-hydroxy-phenyl)-piperazine-1-carboxylic acid tert-butyl ester and1-Methyl-3-pyrrolidinol.

Preparation 45A4-[2-(1-Acetyl-piperidin-3-yloxy)-phenyl]-piperazine-1-carboxylic acidtert-butyl ester

Step 1: 1-(3-Hydroxy-piperidin-1-yl)-ethanone

3-Hydroxypiperidine (3.0 g, 30.0 mmol) was dissolved in THF (40 mL) andmethylene chloride (40 mL). Triethylamine (7.47 g, 74.0 mmol) was added.The mixture was cooled to −78° C. and acetyl chloride (2.35 g, 30.0mmol) was added drop wise. The mixture was warmed to r.t. and stirredunder nitrogen for 1 about an hour. The solvents were removed underreduced pressure, and the resulting oil was purified using silicachromatography (2% methanol in ethyl acetate) to afford the titlecompound (3.57 g, 83%) as an oil.

¹H NMR (CDCl₃) δ 3.90–3.55 (m, 3H), 3.50–3.15 (m, 3H), 2.10 (s, 3H),1.95–1.60 (m, 2H), 1.55–1.25 (m, 2H).

Step 2: The title compound was prepared by using mitsonobu conditionfrom 4-(2-hydroxy-phenyl)-piperazine-1-carboxylic acid tert-butyl esterand 1-(3-Hydroxy-piperidin-1-yl)-ethanone.

¹H NMR (CDCl₃) δ 7.05–6.95 (m, 4H), 3.75–3.70 (m, 3H), 3.60–3.50 (m,4H), 3.35–3.30 (m, 2H), 3.05–3.00 (m, 4H), 1.90–0.185 (m, 2H), 2.00 (s,3H), 1.50 (s, 9H), 1.45–1.40 (m, 2H).

Preparation 46A4-{2-[2-(4-Methanesulfonyl-piperazin-1-yl)-ethoxy]-phenyl}-piperazine-1-carboxylicacid tert-butyl ester

Step 1: 2-(4-Methanesulfonyl-piperazin-1-yl)-ethanol

2-Piperazin-1-yl-ethanol (1.30 g, 1.0 mmol) and triethylamine (2.8 mL, 2mmol) were dissolved in methylene chloride (20 mL). The solution wascooled to 0° C. and methanesulfonyl chloride (0.78 mL, 1 mmol) was addeddropwise. The mixture was warmed to r.t., and the solvent was removedunder reduced pressure. The residue was purified via silica gelchromatography (10% methanol in ethyl acetate) to afford the titlecompound (1.0 g, 48%).

¹H NMR (CDCl₃) δ 3.62–3.75 (m, 2H), 3.23–3.38 (m, 4H), 2.82 (s, 3H),2.57–2.74 (m, 6H). TLC (SiO₂): R_(f)=0.16 (5% methanol in ethyl acetate)

Step 2: The title compound was prepared by using Mitsunobu conditionfrom 4-(2-hydroxy-phenyl)-piperazine-1-carboxylic acid tert-butyl esterand 2-(4-Methanesulfonyl-piperazin-1-yl)-ethanol.

Preparation 47A4-[2-(1-Methanesulfonyl-azetidin-3-yloxy)-phenyl]-piperazine-1-carboxylicacid tert-butyl ester

Step 1: 1-Benzhydryl-3-(2-bromo-phenoxy)-azetidine

Bromophenol (3.8 mL, 32.6 mmol), benzhydrylazetidinol (7.8 g, 32.6 mmol)and triphenylphosphine (12.8 g, 28.9 mmol) were dissolved in THF (200mL) under nitrogen and cooled to 0° C. Diisopropyl azodicarboxylate (9.7mL, 48.9 mmol) was added dropwise over 30 minutes, and the mixture wasstirred for about 96 hours at r.t. Diethyl ether (400 mL) was added, andthe solution was washed with 5 N NaOH (2×50 mL) and brine (50 mL), andthen dried over magnesium sulfate and concentrated to a colorless oil.Purification by silica gel chromatography (4:1 hexanes/ethyl acetate)gave the title compound as a colorless oil (7.0 g, 55%).

¹H NMR (CDCl₃) δ 7.51 (dd, J=7.8, 1.6 Hz, 1H) 7.40–7.43 (m, 4H),7.11–7.33 (m, 8H), 6.79 (td, J=7.7, 1.3 Hz, 1H), 6.58 (dd, 1H, J=8.2,1.3 Hz) 4.78–4.84 (m, 1H,), 4.45 (s, 1H), 3.72–3.77 (m, 2H), 3.13–3.18(m, 2H)

Step 2: 1-[2-(1-Benzhydryl-azetidin-3-yloxy)-phenyl]-piperazine

1-Benzhydryl-3-(2-bromo-phenoxy)-azetidine (3.6 g, 9.16 mmol),piperazine (946 mg, 11 mmol), BINAP (427 mg, 1.38 mmol), Pd₂dba₃ (410mg, 0.46 mmol) and sodium tert-butoxide (1.23 g, 12.8 mmol) weredissolved into toluene (100 mL). The solution was degassed under vacuumand bubbled with nitrogen. The vessel was sealed under nitrogen andheated to 90° C. for about 24 hours. Diethyl ether (500 ml) was added,and the solution was filtered through a bed of celite. The solution wasconcentrated to an orange oil. Purification by silica gel chromatography[1% NH₃ in methanol/ethyl acetate (1 L), 5% NH₃ in methanol/ethylacetate (1 L), and 10% NH₃ in methanol/ethyl acetate (1L)] gave thetitle compound as an orange powder (2.0 g, 65%).

¹H NMR (CDCl₃) δ 7.40–7.43 (m, 4H), 7.25–7.37 (m, 4H), 7.15–7.21 (m,2H), 6.83–6.92 (m, 3H), 6.55 (dd, J=6.5, 1.7 Hz, 11) 4.79–4.87 (m, 1H,),4.44 (s, 1H), 3.71–3.77 (m, 2H), 3.12–3.16 (m, 2H), 3.04 (s, 8H), 1.86(bs, 1H).

Step 3:4-[2-(1-Benzhydryl-azetidin-3-yloxy)-phenyl]-piperazine-1-carboxylicacid tert-butyl ester

4-[2-(1-Benzhydryl-azetidin-3-yloxy)-phenyl]-piperazine-1-carboxylicacid tert-butyl ester (610 mg, 1.53 mmol) and di-tert-butyl dicarbonate(0.883 g, 4.05 mmol) were dissolved into dioxane (3 mL) and water (5 mL)at 0° C. Sodium bicarbonate (340 mg, 4.05 mmol) was added, and themixture was warmed to r.t. and stirred for about 2 hours. Ethyl acetate(100 mL) was added, and the solution was washed with H₂O (3×20 mL),saturated sodium bicarbonate (20 mL)and brine (20 mL) and then driedover magnesium sulfate. Purification by silica gel chromatography(hexanes/ethyl acetate 4:1) gave the title compound as a colorless oil(770 mg, 100%).

¹H NMR (CDCl₃) δ 7.40–7.43 (m, 4H), 7.16–7.34 (m, 6H), 6.85–6.91 (m,3H), 6.55–6.58 (m, 1H), 4.80–4.87 (m, 1H), 4.44 (s, 1H), 3.72–3.77 (m,2H), 3.55–3.59 (m, 4H), 3.13–3.19 (m, 2H), 2.99–3.02 (m, 4H), 1.49 (s,9H). TLC (SiO₂): R_(f)=0.90 (1:1 hexanes/ethyl acetate)

Step 4: 4-[2-(Azetidin-3-yloxy)-phenyl]-piperazine-1-carboxylic acidtert-butyl ester

4-[2-(1-Benzhydryl-azetidin-3-yloxy)-phenyl]-piperazine-1-carboxylicacid tert-butyl ester (750 mg, 1.5 mmol) and ammonium formate (1.8 g, 30mmol) were dissolved into methanol (50 mL) at r.t. The solution wasadded slowly to 10% palladium on carbon (750 mg) in methanol (20 mL)under nitrogen atmosphere. The mixture was stirred for about 48 hoursand filtered through a bed of celite. The solvent was removed underreduced pressure to afford the title compound as a colorless oil (380mg, 60%).

¹H NMR (CDCl₃) δ 6.98–7.04 (m, 3H), 6.72–6.75 (m, 1H), 5.17–5.21 (m,1H,), 4.53 (dd, J=12.4, 6.5 Hz, 2H), 4.18 (dd, J=12.3, 5.0 Hz, 2H),3.57–3.61 (m, 4H), 2.99–3.02 (m, 4H), 1.48 (s, 9H)

Step 5: 4-[2-(Azetidin-3-yloxy)-phenyl]-piperazine-1-carboxylic acidtert-butyl ester (137 mg, 0.41 mmol) and triethylamine (97 μL, 0.7 mmol)were dissolved into THF (2 mL) at 0° C. Mesityl chloride (45 μL, 0.57mmol) was added, and the solution was stirred for about 16 hours. Thesolution was concentrated and filtered through a silica gel plug with1:1 hexanes/ethyl acetate. The final compound was isolated as acolorless oil (120 mg, 71%)

¹H NMR (CDCl₃) δ 6.90–7.10 (m, 3H), 6.65–6.75 (m, 1H), 4.95–5.01 (m,1H,), 4.25–4.35 (m, 2H), 4.05–4.15 (m, 2H), 3.55–3.65 (m, 4H), 2.97–3.02(m, 4H), 2.91 (s, 3H), 1.48 (s, 9H). TLC (SiO₂): R_(f)=0.38 (1:1hexanes/ethyl acetate)

Preparation 48A4-[2-(1-Methyl-azetidin-3-yloxy)-phenyl]-piperazine-1-carboxylic acidtert-butyl ester

4-[2-(Azetidin-3-yloxy)-phenyl]-piperazine-1-carboxylic acid tert-butylester (Preparation 47A, Step 4) (140 mg, 0.42 mmol) was dissolved inCH₂Cl₂ (2 mL). Acetic acid (0.15 mL) and formalin (85 μL, 1.05 mmol)were added, and the mixture was cooled to 0° C. Sodiumtriacetoxyborohydride (249 mg, 1.17 mmol) was added, and the solutionwas stirred for about 16 hours. Methylene chloride (15 mL) was added,and the solution was washed with saturated NaHCO₃ (5 mL) and brine (5mL) and then dried over magnesium sulfate and concentrated to acolorless oil (300 mg, 21%).

¹H NMR (CDCl₃) δ 6.91–6.98 (m, 3H), 6.58–6.61 (m, 1H), 4.72–4.79 (m,1H), 3.83–3.88 (m, 2H), 3.60 (t, J=5.1 Hz, 2H), 3.12–3.17 (m, 2H), 3.01(t, J=5.1 Hz, 4H), 2.72 (s, 3H), 1.48 (s, 9H)

Preparation 49A4-{2-[1-(Propane-2-sulfonyl)-piperidin-3-yloxy]-phenyl}-piperazine-1-carboxylicacid tert-butyl ester

Step 1:4-[2-(1-Benzyloxycarbonyl-piperidin-3-yloxy)-phenyl]-piperazine-1-carboxylicacid tert-butyl ester

(S)-3-Hydroxy-piperidine-1-carboxylic acid benzyl ester (2.34 g, 8.6mmol) was dissolved in THF (40 mL) at room temperature.4-(2-Hydroxy-phenyl)-piperazine-1-carboxylic acid tert-butyl ester (2.4g, 8.6 mmol) and triphenylphosphine (2.7 g, 10.32 mmol) were added.Diisopropyl azodicarboxylate (2.05 mL, 10.32 mmol) was added dropwise tothe solution. The reaction was stirred overnight, concentrated underreduced pressure, and purified using silica gel chromatography (100%ethyl acetate) to afford the title compound (2.5 g, 55%).

¹H NMR (CDCl₃) δ 7.30–7.20 (m, 5H), 6.65–6.50 (m; 4H), 5.35 (s, 2H),3.75–3.70 (m, 3H), 3.65–3.60 (m, 4H), 3.35–3.30 (m, 2H), 3.25–3.20 (m,4H), 1.85–1.60 (m, 4H), 1.50 (s, 9H)

Step 2:4-{2-[1-(Propane-2-sulfonyl)-piperidin-3-yloxy]-phenyl}-piperazine-1-carboxylicacid tert-butyl ester

4-[2-(1-Benzyloxycarbonyl-piperidin-3-yloxy)-phenyl]-piperazine-1-carboxylicacid tert-butyl ester (2.5 g, 4.7 mmol) was dissolved in methanol (20mL). Palladium on carbon (50% water, 2.1 g) was added. A hydrogenatmosphere was established using balloons, and the mixture was stirredat r.t. for about 4 hours. The catalyst was removed by filtration withethyl acetate through a bed of celite. The compound was isolated byrunning a silica gel plug with a gradient of 100% ethyl acetate (200 mL)to triethylamine in ethyl acetate (10%). The isolated product (0.382 g,0.96 mmol) was dissolved in methylene chloride (10 mL) at 0° C., andtriethylamine (0.4 mL, 2.9 mmol) was added. Isopropylsulfonyl chloride(0.13 mL, 1.15 mmol) was added dropwise, and the mixture was warmed tor.t. for about 30 minutes, which was then concentrated under reducedpressure and purified using silica gel chromatography (75% ethyl acetatein hexanes) to afford the final compound (0.39 g, 80%).

¹H NMR (CDCl₃) δ 7.25–6.85 (m, 4H), 3.55–3.50 (m, 4H), 3.05–2.95 (m,4H), 2.15–1.60 (m, 4H), 1.50 (s, 9H), 1.45–1.40 (m, 6H), 1.30–1.25 (m,5H)

Preparation 50A4-[2-(1-Benzoyl-piperidin-3-yloxy)-phenyl]-piperazine-1-carboxylic acidtert-butyl ester

4-[2-(Piperidin-3-yloxy)-phenyl]-piperazine-1-carboxylic acid tert-butylester (Preparation 49A) (311 mg, 0.86 mmol) was dissolved in CH₂Cl₂ (15mL) under nitrogen and cooled to 0° C. Benzoyl chloride (0.3 mL, 2.58mmol) was added followed by dropwise addition of triethylamine (0.48 mL,3.44 mmol). The mixture was warmed to r.t. and stirred for about 16hours. Methylene chloride (20 mL) was added, and the solution was washedwith saturated NaHCO₃ (20 mL)and brine (20 mL), and then dried overmagnesium sulfate. The solvents were removed under reduced pressure, andthe residue was purified by flash column chromatography on silica toafford the final compound as a white solid (330 mg, 83%).

¹H NMR (CDCl₃) δ 7.25–7.45 (m, 5H), 6.90–7.10 (m, 3H), 6.82–6.90 (m,1H), 4.15–4.30 (m, 1H), 3.42–3.85 (m, 8H), 2.86–3.20 (m, 4H), 1.85–2.10(m, 3H), 1.51–1.69 (m, 1H), 1.48 (s, 9H) TLC (SiO₂): R_(f)=0.2 (1:1ethyl acetate/hexanes)

Preparation 51A4-[2-(1-Isobutyryl-piperidin-3-yloxy)-phenyl]-piperazine-1-carboxylicacid tert-butyl ester

4-[2-(Piperidin-3-yloxy)-phenyl]-piperazine-1-carboxylic acid tert-butylester (Preparation 49A, Step 2) (340 mg, 0.94 mmol) was dissolved inCH₂Cl₂ (15 mL) under nitrogen and cooled to 0° C. Isobutyryl chloride(0.29 mL, 2.82 mmol) was added followed by dropwise addition oftriethylamine (0.52 mL, 3.76 mmol). The mixture was warmed to r.t. andstirred for about 16 hours. Methylene chloride (20 mL) was added, andthe solution was washed with saturated NaHCO₃ (20 mL)and brine (20 mL),and then dried over magnesium sulfate. The solvents were removed underreduced pressure, and the residue was purified by flash columnchromatography on silica gel, eluting with ethyl acetate/hexanes (1:1),to afford the final compound as a white solid (322 mg, 80%). TLC (SiO₂):R_(f)=0.42 (1:1 ethyl acetate/hexanes)

Preparation 52A4-[2-(1-Benzenesulfonyl-piperidin-3-yloxy)-phenyl]-piperazine-1-carboxylicacid tert-butyl ester

Step 1:4-[2-(1-Benzyloxycarbonyl-piperidin-3-yloxy)-phenyl]-piperazine-1-carboxylicacid tert-butyl ester

3-Hydroxy-piperidine-1-carboxylic acid benzyl ester (2.34 g, 8.6 mmol)was dissolved in THF (40 mL) at r.t.4-(2-Hydroxy-phenyl)-piperazine-1-carboxylic acid tert-butyl ester (2.4g, 8.6 mmol) and triphenylphosphine (2.7 g, 10.32 mmol) were added.Diisopropyl azodicarboxylate (2.05 mL, 10.32 mmol) was added dropwise tothe solution. The mixture was stirred overnight, and then concentratedunder reduced pressure and purified using silica gel chromatography(100% ethyl acetate) to afford the title compound (2.5 g, 55%).

¹H NMR (CDCl₃) δ 7.30–7.20 (m, 5H), 6.65–6.50 (m, 4H), 5.35 (s, 2H),3.75–3.70 (m, 3H), 3.65–3.60 (m, 4H), 3.35–3.30 (m, 2H), 3.25–3.20 (m,4H), 1.85–1.60 (m, 4H), 1.50 (s, 9H)

Step 2: The compound of Step 1 (2.5 g, 4.7 mmol) was dissolved inmethanol (20 mL). Palladium on carbon (50% water, 2.1 g) was added. Ahydrogen atmosphere was established using balloons, and the mixture wasstirred at r.t. for about 4 hours. The catalyst was removed byfiltration with ethyl acetate through a bed of celite. The compound wasisolated by running scrub-plug with a gradient of 100% ethyl acetate(200 mL) to triethylamine in ethyl acetate (10%). The isolated product(0.36 g, 0.9 mmol) was dissolved in methylene chloride (10 mL) at 0° C.,and triethylamine (0.38 mL, 2.7 mmol) was added. Benzenesulfonylchloride (0.15 mL, 1.1 mmol) was added drop wise to the reactionmixture. The solution warmed to r.t., concentrated under reducedpressure and purified using silica gel chromatography (75% ethyl acetatein hexanes) to afford the final compound (0.400 g, 82%).

¹H NMR (CDCl₃) δ 7.95–7.85 (m, 2H), 7.55–7.30 (m, 3H), 6.65–6.50 (m,4H), 3.75–3.70 (m, 1H), 3.65–3.60 (m, 4H), 3.25–3.20 (m, 4H), 3.15–3.10(m, 2H), 2.75–2.70 (m, 2H), 1.90–1.80 (m, 2H), 1.65–1.55 (m, 2H), 1.50(s, 9H)

Preparation 53A

Step 1:4-[2-(1-benzyl-piperidin-3-yloxy)-phenyl]-piperazine-1-carboxylic acidtert-butyl ester was s prepared by using Mitsunobu condition from4-(2-hydroxy-phenyl)-piperazine-1-carboxylic acid tert-butyl ester and(S)-1-benzyl-piperidin-3-ol.

Step 2: 4-[2-(Piperidin-3-yloxy)-phenyl]-piperazine-1-carboxylic Acidtert-butyl ester

A solution of the compound from Step 1 (2.07 g, 4.59 mmol) in ethanol(50 mL) was treated with palladium on carbon (50% by weight with water,1.0 g), and the mixture was shaken on a Parr hydrogenation apparatus forabout 48 hours. The solution was filtered through celite and, thesolvent was removed under reduced pressure to yield the title compoundas a clear oil (1.10 g, 66%).

¹H NMR (CD₃OD) δ 6.93–7.10 (m, 4H), 4.35–4.47 (m, 1H), 4.52–4.68 (m,5H), 3.03–3.18 (m, 2H), 2.64–3.02 (m, 6H), 1.77–2.15 (m, 3H), 1.53–1.66(m, 1H), 1.48 (s, 9H)

Step 3: Triethylamine (0.26 g, 2.53 mmol) was added to a solution of thecompound from Step 2 (0.37 g, 1.01 mmol) in methylene chloride (10mL)/THF (10 mL) at −78° C. under nitrogen. Methanesulfonyl chloride wasadded dropwise, and the reaction was allowed to warm to r.t. and stirredfor about 2 hours. The mixture was diluted with diethyl ether (200 mL),filtered, and the solvent was removed under reduced pressure. Theresidue was purified by flash column chromatography on silica gel byeluting with ethyl acetate/hexanes (1:1) to afford the final compound asa clear oil (0.26 g, 56%).

¹H NMR (CDCl₃) δ 6.82–7.05 (m, 4H), 4.34–4.50 (m, 1H), 3.73–3.89 (m,1H), 3.45–3.67 (m, 5H), 2.87–3.24 (m, 8H), 2.03–2.21 (m, 1H), 1.87–2.01(m, 1H), 1.64–1.72 (m, 2H), 1.55 (m, 3H), 1.52 (s, 9H)

Preparation 54A4-[2-(1-Propionyl-piperidin-3-yloxy)-phenyl]-piperidine-1-carboxylicAcid tert-Butyl Ester

Propionyl chloride (0.11 g, 1.21 mmol) was added dropwise to a solutionof 4-[2-(piperidin-3-yloxy)-phenyl]-piperazine-1-carboxylic acidtert-butyl ester (Preparation 53A) (0.37 g, 1.01 mmol) and DIPEA (0.20g, 1.52 mmol) in methylene chloride (20 mL) at 0° C. under nitrogen. Themixture was warmed to r.t. and stirred overnight. The solution wasdiluted with ethyl acetate (400 mL) and washed with water (45 mL),saturated aqueous sodium bicarbonate (45 mL) and brine (45 mL), whichwas then dried over anhydrous sodium sulfate. The solvent was removed,and the residue was purified by flash column chromatography on silicagel by eluting with ethyl acetate/hexanes (1:1) to provide the finalcompound as a clear oil (0.26 g, 61%).

¹H NMR (CDCl₃) δ 6.81–7.02 (m, 4H), 4.20–4.52 (m, 2H), 3.45–3.65 (m,6H), 3.25–3.42 (m, 1H), 2.86–3.12 (m, 4H), 2.30–2.47 (m, 1H), 2.08–2.25(m, 1H), 1.77–2.03 (m, 2H), 1.61 (s, 2H), 1.51 (s, 9H), 1.01–1.23 (m,3H)

Preparation 55A4-[2-(2-Methanesulfonylamino-propoxy)-phenyl]-piperazine-1-carboxylicacid tert-butyl ester

Step 1: 4-[2-(2-Amino-propoxy)-phenyl]-piperazine-1-carboxylic acidtert-butyl ester

4-[2-(2-Benzyloxycarbonylamino-propoxy)-phenyl]-piperazine-1-carboxylicacid tert-butyl ester (5.7 g, 12.15 mmol) and 10% wet palladium oncarbon (5.0 g) were dissolved in methanol (200 mL) at r.t. The solutionwas stirred under hydrogen for about 4 hours. The mixture was filteredthrough a bed of celite. The solvent was removed under reduced pressureto afford the title compound as a colorless oil (4.2 g, 100%).

¹H NMR (CDCl₃) δ 6.89–7.05 (m, 4H), 4.01 (dd, J=9.6, 4.1 Hz, 1H), 3.86(t, J=7.8 Hz, 1H), 3.56–3.86 (m, 6H), 3.36–3.45 (m, 1H), 1.47 (s, 9H),1.25 (d, J=6.5 Hz, 3H)

Step 2: The compound of Step 1 (750 mg, 2.24 mmol) and triethylamine(0.52 mL, 3.7 mmol) were dissolved in THF (10 mL) at 0° C. Mesitylchloride (0.24 mL, 3.14 mmol) was added, and the solution was stirredfor about 16 hours. Ethyl acetate (50 mL) was added, and the solutionwas washed with saturated NaHCO₃ (2×10 mL) and brine (10 mL) and thendried over magnesium sulfate. The solution was concentrated to affordthe final compound as a colorless oil (930 mg, 100%).

¹H NMR (CDCl₃) δ 6.91–7.01 (m, 4H), 6.27 (d, J=7.9 Hz, 1H), 4.01–4.19(m, 2H), 3.77–3.86 (m, 1H), 3.63 (s, 4H), 3.09–3.16 (m, 2H), 2.87–2.94(m, 2H), 2.82 (s, 3H), 1.48 (s, 9H), 1.41 (d, J=6.9 Hz, 3H). TLC (SiO₂):R_(f)=0.80 (3:1 ethyl acetate/hexanes)

Preparation 56A4-{2-[2-(Methanesulfonyl-methyl-amino)-propoxy]-phenyl}-piperazine-1-carboxylicacid tert-butyl ester

4-[2-(2-Methanesulfonylamino-propoxy)-phenyl]-piperazine-1-carboxylicacid tert-butyl ester (470 mg, 1.14 mmol) was dissolved in THF (20 mL)and cooled to 0° C. Sodium hydride (29 mg of 60% dispersion in oil, 1.2mmol) was added followed by dropwise addition of methyl iodide (78 μL,1.25 mmol). The mixture was warmed to r.t. and stirred for about 36hours. Ethyl acetate (20 mL) was added, and the solution was washed withsaturated NaHCO₃ (5 mL), H₂O (5 mL) and brine (5 mL) and then dried overmagnesium sulfate. The solvents were removed under reduced pressure toafford the final compound as a clear oil (486 mg, 100%).

¹H NMR (CDCl₃) δ 6.85–7.03 (m, 4H), 4.06–4.15 (m, ¹H,), 3.89 (dd,J=10.0, 4.5 Hz, 1H), 3.50–3.62 (m, 4H), 2.85–3.05 (m, 4H), 2.98 (s, 3H),2.85–3.05 (m, 4H), 2.88 (s, 3H), 1.48 (s, 9H), 1.32 (d, J=7.0 Hz, 3H)

Preparation 57A4-[2-(2-Piperidin-1-yl-propoxy)-phenyl]-piperazine-1-carboxylic acidtert-butyl ester

Step 1:4-[2-(2-Benzyloxycarbonylamino-propoxy)-phenyl]-piperazine-1-carboxylicacid tert-butyl ester

(3-Hydroxy-2-methyl-propyl)-carbamic acid benzyl ester (5.0 g, 24.0mmol) was dissolved in THF (175 mL) at 0° C.4-(2-hydroxy-phenyl)-piperazine-1-carboxylic acid tert-butyl ester (6.7g, 24.0 mmol) and triphenylphosphine (7.6 g, 29.0 mmol) were added.Diisopropyl azodicarboxylate (5.75 mL, 29.0 mmol) was added dropwise,and the mixture was stirred overnight. The mixture was concentratedunder reduced pressure and purified using silica gel chromatography (25%ethyl acetate in hexanes) to afford the title compound (5.33 g, 47%).

¹H NMR (CDCl₃) δ 7.30–7.20 (m, 5H), 7.00–6.80 (m, 4H), 5.75 (s, 1H),5.05 (s, 2H), 4.10–4.00 (m, 3H), 3.95–3.90 (m, 1H), 3.55 (s, 4H),3.10–3.00 (m, 2H), 2.90–2.80 (m, 2H), 2.05 (s, 1H)

Step 2: The compound of Step 1 (5.33 g, 11.36 mmol) was dissolved inmethanol (200 mL), and palladium on carbon (50% water, 5.0 g) was thenadded. A hydrogen atmosphere was established using balloons and thereaction stirred at r.t. for about 4 hours. The catalyst was removed byfiltration with ethyl acetate (200 mL) through a bed of celite. Compoundwas isolated, and the residue (1.0 g, 2.13 mmol) was dissolved inacetonitrile (55 mL). Potassium carbonate (1.18 g, 8.52 mmol) was added,and the mixture was heated to reflux and then diiodopentane (0.76 g,2.34 mmol) in acetonitrile (10 mL) was added dropwise. The reaction wasstirred overnight and cooled to r.t., filtered and concentrated underreduced pressure. The compound was purified by using silica gelchromatography (1% ammonium hydroxide in ethyl acetate) to afford thefinal compound (0.95 g, 83%).

¹H NMR (CDCl₃) δ 6.95–6.80 (m, 4H), 4.10–4.00 (m, 1H), 3.85–3.80 (m,1H), 3.60–3.55 (m, 4H), 3.35–3.30 (m, 1H), 3.00 (s, 4H), 2.75–2.50 (m,4H), 1.50 (s, 9H), 1.45–1.30 (m, 6H), 1.10 (s, 3H)

Preparation 58A4-[2-(2-Diethylamino-propoxy)-phenyl]-piperazine-1-carboxylic acidtert-butyl ester

4-[2-(2-Amino-propoxy)-phenyl]-piperazine-1-carboxylic acid tert-butylester (0.35 g, 0.75 mmol) was dissolved in methanol (25 mL), and thereaction was cooled to 0° C. Acetaldehyde (0.363 g, 8.25 mmol) andsodium triacetoxyborohydride (0.795 g, 3.75 mmol) were added. A refluxcondenser was fitted and the flask was stirred at r.t. for about 24hours. The solution was extracted with diethyl ether (3×50 mL) and thenconcentrated under reduced pressure to afford the final compound (0.360g, 90%).

¹H NMR (CDCl₃) δ 6.95–6.80 (m, 4H), 4.10–4.00 (m, 1H), 3.85–3.80 (m,1H), 3.60–3.55 (m, 4H), 3.35–3.30 (m, 1H), 3.00 (s, 4H), 2.75–2.50 (m,4H), 1.50 (s, 9H), 1.20–1.15 (m, 4H), 1.10–1.00 (m, 5H)

Preparation 59A 4-[2-(N-methyl-piperidin-4-yloxy)-phenyl]-piperazine

Step 1: N-Cbz-4-[2-(N-boc-piperidin-4-yloxy)-phenyl]-piperazine

To a solution of 4-[2-(N-boc-piperidin-4-yloxy)-phenyl]-piperazine (1.5g, 4.1 mol, 1.0 eq.) in DCM (20 mL) was added Et₃N (0.8 mL, 5.74 mmol,1.4 eq.), di-carbobenzoyloxy anhydride (1.4 g, 4.9 mmol, 1.2 eq.) andDMAP (25 mg, 0.2 mmol, 0.5 eq.). The mixture was stirred at r.t.overnight. The solution was diluted with DCM (100 mL) and washed withsaturated aqueous NaHCO₃ (50 mL) and brine (50 mL). The organic layerwas dried (Na₂SO₄), filtered and concentrated to afford the titlecompound (2.03 g, 4.1 mmol, 100%).

LRMS (electrospray): 496.4 (M+1).

Step 2: N-Cbz-4-[2-(piperidin-4-yloxy)-phenyl]-piperazine

To a solution of Step 2 (2.03 g, 4.1 mmol) in CH₂Cl₂ (10 mL) and DMS (2mL) was added TFA (10 mL). After stirring for about 2 hours, thesolution was diluted with heptane and concentrated (2×). The residuewere dissolved in CH₂Cl₂ and washed with saturated aqueous sodiumbicarbonate. The aqueous solution was extracted with CH₂Cl₂ (3×). Thecombined organic extracts were dried (Na₂SO₄), filtered andconcentrated. The product was purified by SCX (10 g) ion exchangechromatography. Further purification by flash chromatography (125 gSiO₂, linear gradient, 40 mL/min, 0%–10% 2.0M NH₃ in MeOH/CH₂Cl₂ for 20minutes and then 10% 2.0M NH₃ in MeOH/CH₂Cl₂ for 46 minutes) affordedtitle compound (1.05 g, 2.65 mmol, 65%). LRMS (electrospray): 396.3(M+1).

Step 3: To a solution of the compound obtained from Step 2 (1.0 g, 2.5mmol, 1.0 eq.) in EtOH (8 mL) was added formaldehyde (37 wt % in H₂O,1.0 mL, 12.5 mmol, 5.0 eq.) and formic acid (0.5 mL, 12.5 mmol, 5.0eq.). The mixture was heated at 70° C. and stirred at that temperatureovernight. The mixture was cooled to r.t. and concentrated. Purificationby SCX (10 g) ion-exchange chromatography afforded Cbz-protected titlecompound (1.0 g, 2.44 mmol, 98%). LRMS (electrospray): 410.1(M+1).

Step 4: To a solution of Cbz-protected title compound made above (480mg, 1.17 mmol, 1.0 eq.) in IPA (15 mL) was added 10% Palladium on carbon(96 mg, 20 wt %). The reaction mixture was stirred at r.t. under H₂ (1atm) over night. The reaction mixture was filtered through a pad ofcelite and concentrated to afford final compound (315 mg, 1.14 mmol,98%). LRMS (electrospray): 276.1 (M+1).

Preparation 60A Dimethyl-[2-(2-piperazin-1-yl-phenoxy)-ethyl]-amine

To a solution of 1-Boc-4(2-hydroxyphenyl)-piperazine (300 mg, 1.08mmol), 2-dimethylaminoethyl chloride hydrochloride (233 mg, 1.62 mmol),K₂CO₃ (450 mg, 3.26 mmol), and KI (357 mg, 2.15 mmol) in DMF (10 mL) wasadded 18-crown-6 (1.42 g, 5.37 mmol). After stirring overnight, waterwas added and the solution extracted with CH₂Cl₂ (3×). The combinedorganic extracts were concentrated to an oil. The oil was loaded onto a10 g SCX ion exchange column equilibrated with MeOH. The column wasflushed with 20 mL of MeOH, 20 mL of 0.2 M NH₃ in MeOH, and 20 mL of 2 MNH₃ in MeOH. The fractions containing desired product were combined andconcentrated to an oil. To a solution of the oil in MeOH (2 mL) wasadded 1M HCl (6 mL) in Et₂O. After stirring overnight the solution wasconcentrated to an oil to obtain the final compound. MS: 250.2 (M+1).

Preparation 61AN-boc-4-{2-[2-(ethyl-methanesulfonyl-amino)-2-methyl-propoxy]-phenyl}-piperazine

Step 1: To a solution ofN-boc-1-[2-(2-methyl-2-nitro-propoxy)-phenyl]-piperazine (1.22 g) in 50mL of DMF was added K₂CO₃ (3.5 g, 25 mmol). After stirring for about anhour, 2-methyl-2-nitropropyl-p-toluenesulfonate (3 g, 11 mmol) wasadded. After stirring overnight at 100° C., the solution was cooled tor.t. and diluted with EtOAc. The mixture was washed with water (2×) andbrine, and then dried (Na₂SO₄), filtered and concentrated. Purificationby silica gel chromatography (4×20 cm SiO₂, 10 to 20% EtOAc/hexanes,over 45 minutes at 35 mL/min) afforded about 2.43 g (8.86 mmol, 89%) of1-bromo-2-(2-methyl-2-nitro-propoxy)-benzene as a white solid. GCMS(EI): 273 [M].

Step 2: A solution of 1-bromo-2-(2-methyl-2-nitro-propoxy)-benzene (2.30g, 8.4 mmol), piperazine (1.8 g, 20 mmol), Pd₂dba₂ (384 mg, 0.4 mmol),BINAP (784 mg, 1.26 mmol), and NaOtBu (1.13 g, 12 mmol) in 34 mL oftoluene was heated to 90° C. for 5 hrs. The solution was concentrated,dissolved in CH₂Cl₂ and filtered through celite. Purification by silicagel chromatography (4×20 cm SiO₂, 0,1,2,3,4,5,7,9,11% 2M NH₃ inMeOH/CH₂Cl₂ step gradient, 12 min each at 35 mL/min) afforded 1.72 g(6.15 mmol, 73%) of 1-[2-(2-methyl-2-nitro-propoxy)-phenyl]-piperazineas a yellow oil. LRMS (electrospray): 280.1 [M+1].

Step 3: To a solution of compound from Step 2 (1.68 g, 6.01 mmol), DMAP(40 mg, 0.33 mmol), Et₃N (1.84 mL, 13.2 mmol) in 60 mL of CH₂Cl₂ wasadded Boc₂O (1.44 g, 6.60 mmol). After stirring overnight, the solutionwas washed with 1 M HCl, water, saturated aqueous sodium bicarbonate andbrine, and then dried (Na₂SO₄), filtered and concentrated to affordabout 2.0 g (5.28 mmol, 88%)N-boc-1-[2-(2-methyl-2-nitro-propoxy)-phenyl]-piperazine as a yellowoil. LRMS (electrospray): 380.2 [M+1].

Step 4: To a solution ofN-boc-1-[2-(2-methyl-2-nitro-propoxy)-phenyl]-piperazine (1.22 g, 3.22mmol) and ammonium formate (2.0 g, 32 mmol) in 15 mL of MeOH and 5 mL ofTHF was added 10% Pd/c (500 mg). After stirring for about 3 days under60 psi of H₂, the solution was filtered through celite and concentrated.The residue was partitioned between water and EtOAc. The aqueoussolution was extracted with EtOAc (3×). The combined organic solutionswere washed with brine and then dried (Na₂SO₄), filtered andconcentrated to afford about 1.17 g (3.35 mmol, 100%) ofN-boc-4-[2-(2-amino-2-methyl-propoxy)-phenyl]-piperazine as an off whitesolid. LRMS (electrospray): 350.2 [M+1].

Step 5: To a solution of compound from Step 4 (175 mg, 0.5 mmol) andEt₃N (210 uL, 1.5 mmol) in 5 mL of CH₂Cl₂ at 0° C. was added MsCl (41uL, 0.53 mmol) at 10 uL increment until starting material was no longerpresent by TLC. The mixture was quenched with saturated aqueous sodiumbicarbonate, diluted with EtOAc and washed with 1 M HCl, water andbrine, which was then dried (Na₂SO₄), filtered and concentrated toafford about 155 mg (0.36 mmol, 73%) ofN-boc-4-[2-(2-methanesulfonylamino-2-methyl-propoxy)-phenyl]-piperazineas a yellow solid. LRMS (electrospray): 428.1 [M+1].

Step 6: To a solution ofN-boc-4-[2-(2-methanesulfonylamino-2-methyl-propoxy)-phenyl]-piperazine(197 mg, 0.46 mmol) in 4 mL of DMF was added NaH (55 mg of a 60%dispersion in oil, 1.38 mmol). After stirring for about 30 minutes, EtI(550 uL, 6.88 mmol) was added. After stirring at 45° C. for about anhour, the mixture was quenched with saturated aqueous sodiumbicarbonate. The mixture was diluted with EtOAc, washed with water andbrine, and then dried (Na₂SO₄), filtered and concentrated. The materialwas combined with 0.36 mmol of crude material from a previous reactionand purified by silica gel chromatography (35 g SiO₂, 10 to 30%EtOAc/hexanes, over 30 minutes at 35 mL/min.) to afford about 323 mg(0.71 mmol, 86% combined yield) of the final compound. LRMS(electrospray): 456.2 [M+1]

Preparation 62A 4-[2-(2-Morpholin-4-yl-ethoxy)-phenyl]-piperazine

To a solution of 2-bromophenol (50 g, 0.289 mol), K₂CO₃ (175 g, 1.27mol) and DMF (1500 mL) was added chloroethyl morpholine-HCl (59 g,0.317). After stirring for about 4 hours at 100° C., 2-bromophenol (4 g,0.023 mol) was added. After stirring for about 3 hours at 100° C., thesolution was cooled to r.t. and diluted with 4L of EtOAc. The solutionwas washed with water, 5 M NaOH, water and brine, and then dried(Na₂SO₄), filtered, and concentrated to afford about 78 g (0.273 mol,94%) of a colorless oil. The product was coupled with piperazine underBuchwald condition to afford the final compound. LRMS (electrospray):292.29 [M+1]

Preparation 63A 1-[2-(2-Morpholin-4-yl-ethoxy)-phenyl]-[1,4]diazepan

The title compound was prepared in a manner similar to Preparation 63Aexcept that 4-[2-(2-bromo-phenoxy)-ethyl]morpholine was coupled tohomopiperazine.

Preparation 64A4-(2-Diethylcarbamoylmethoxy-phenyl)-piperazine-1-carboxylic acidtert-butyl ester

4-(2-hydroxy-phenyl)-piperazine-1-carboxylic acid tert-butyl ester (500mg, 1.8 mmol) was dissolved in DMF (2 mL), and sodium hydride (1.98mmol) was added. The mixture was stirred for about 10 minutes and2-chloro-N,N-diethyl-acetamide (269 mg, 1.8 mmol) was added. The mixturewas heated to 80° C. for about 2 hours. The mixture was concentrated anddeprotected, which was then chromatographed to give about 395 mg of thefinal compound as a clear oil (75%). MS found 292.2

Preparation 65A2,2,2-Trifluoro-1-[3-(2-piperidin-4-yl-phenoxy)-S-pyrrolidin-1-yl]-ethanone

Step 1:4-(4,4,5,5-Tetramethyl-[1,3,2]dioxoborolan-2-yl)-3,6-dihydro-2H-pyridine-1-carboxylicacid tert-butyl ester

4-Trifluoromethanesulfonyloxy-3,6-dihydro-2H-pyridine-1-carboxylic acidtert-butyl ester [Synthesis 1991 (11), 993–995, 8.2 g, 24.7 mmol],bis(pinacolat)diboran(6.93 g, 27.3 mmol),dichloro(1,1-bisdiphenyl-phosphino(ferrocene)Pd(II) dichloromethaneadduct (0.54 g, 0.738 mmol), 1,1′-bis(diphenylphosphino)ferrocene (0.410g, 0.738 mmol) and potassium acetate (6.6 g, 66.8 mmol) were mixed indioxane (150 ml) and degassed. The mixture was heated to 80° C. forabout 4 hours and then stirred at r.t. overnight. The mixture wasdiluted with CH₂Cl₂, filtered through celite, and concentrated to anoil. Oil was chromatographed on silica by eluting with 1:3ethylacetate/hexane to afford the title compound (6.5 g). MS: 310 (M+1)

Step 2:4-{2-[1-(2,2,2-Trifluoro-acetyl)-S-pyrrolidin-3-yloxy]phenyl}-3,6-dihydro-2H-pyridine-1-carboxylicacid tert-butyl ester

1-[3-(2-bromo-phenoxy)-S-pyrrolidin-1-yl]-2,2,2-tritluoro-ethanone (1.19g, 3.52 mmol), the compound of Step 1 (1.31 g, 4.22 mmol), potassiumcarbonate (1.46 g, 10.56 mmol), and dichloro(1,1′-bis(diphenylphosphino) ferrocene) palladium (II) dichloromethane adduct(154 mg, 0.22 mmol) were combined and placed in DMF (12 mL), degassedand heated at 90° C. for about 16 hours. The mixture was cooled to r.t.and diluted with DCM (100 mL). The mixture was filtered over celite andDCM was removed by vacuum. The mixture was partitioned between 500 mL1/1 EtOAc/hexane and 300 mL water, and the aqueous phase was washed with(200 mL) of the 1/1 EtOAc/hexane. The organic phase was washed withwater (200 mL), concentrated and chromatographed on silica gel to affordthe product (486 mg, 31%) as a yellow oil.

Step 3:N-Boc-2,2,2-Trifluoro-1-[3-(2-piperidin-4-yl-phenoxy)-S-pyrrolidin-1-yl]-ethanone

The compound of Step 2 (486 mg, 1.10 mmol) was added to a slurry of Pt/C10% (500 mg) in the solvent system (EtOAc/IPA 50 mL). The mixture waspressurized to 50 psi hydrogen for 2 days. The mixture was filteredthrough celite and concentrated to give a clear oil (467 mg, 96%).

MS found 343.2

Step 4: The compound of Step 3 (467 mg, 1.06 mmol) was placed in thesolvent system (TFA/DCM 1/1 10 mL) and stirred for about 3 hours at r.t.The mixture was quenched with sodium bicarbonate solution and extracted,and the organic phase was concentrated to afford about 286 mg of thefinal compound (84%).

MS found 343.2 (M+1)

Preparation 66A 4-[2-(2-Piperidin4-ylphenoxy)-ethyl]-morpholine

The title compound was prepared by following a substantially similarprocedure as described in Preparation 65A except that4-(4,4,5,5-Tetramethyl-[1,3,2]dioxoborolan-2-yl)-3,6-dihydro-2H-pyridine-1-carboxylicacid tert-butyl ester was coupled with1-[3-(2-bromo-phenoxy)-morpholin-1-yl]-ethanone. MS found 291.2

Preparation 67A2,2,2-Trifluoro-1-[3-(2-R-piperidin-4-yl-phenoxy)-piperidin-1-yl]-ethanone

The title compound was prepared by following a substantially similarprocedure as described in Preparation 65A except that4-(4,4,5,5-Tetramethyl-[1,3,2]dixoaborolan-2-yl)-3,6-dihydro-2H-pyridine-1-carboxylicacid tert-butyl ester was coupled with1-[3-(2-Iodo-phenoxy)-1-R-methyl-piperidin-1-yl]-2,2,2-trifluoro-ethanone.

MS found 357.1

Preparation 68A 1-[2-(1-Methyl-S-piperidin-3-yloxy)-phenyl-piperidine

The title compound was prepared by following a substantially similarprocedure as described in Preparation 65A except that4-(4,4,5,5-Tetramethyl-[1,3,2]dioxoborolan-2-yl)-3,6-dihydro-2H-pyridine-1-carboxylicacid tert-butyl ester was coupled with3-(2-Bromo-phenoxy-1-S-methyl-piperidine. MS found 375.2

Preparation 69ADiethyl-[1-methyl-2-(2-piperidin-4-yl-phenoxy)-ethyl]-amine

The title compound was prepared by following a substantially similarprocedure as described in Preparation 65A except that4(4,4,5,5-Tetramethyl-[1,3,2]dixoaborolan-2-yl)-3,6-dihydro-2H-pyridine-1-carboxylicacid tert-butyl ester was coupledwith[2-(2-Bromo-phenoxy)-1-methyl-ethyl]-diethyl-amine. MS 291 (M+1)

C Domain Preparations:

The protected amino acid derivatives corresponding to the B and Cdomains are, in many cases, commercially available. Other protectedamino acid derivatives can be prepared by following known literaturemethod (See Williams, R. M. Synthesis of Optically Active α-Amino Acids,Pergamon Press: Oxford, 1989). The following provides the preparation ofC domains.

Preparation 1C1-Methoxycarbonylmethyl-1,3-dihydro-isoindole-2-carboxylic acidtert-butyl ester

Step A: (2-Bromo-benzyl)-carbamic acid tert-butyl ester

To a mixture of 125.0 g (561.8 mmol) of 2-bromobenzylamine hydrochlorideand 170.7 g (1236.0 mmol) of potassium carbonate in 300 mL of 50%THF/water was added 134.9 g (618.0 mmol) of di-tert-butyl dicarbonate infour portions over 20 minutes. The mixture was stirred at r.t. for about16 hours and then diluted with 300 mL of ethyl acetate and 300 mL ofwater. The organic portion was separated and the aqueous portion wasextracted three times with 200 mL each of ethyl acetate. The combinedethyl acetate portions were washed once with 250 mL of 10% aqueoussodium bisulfate. The organic portion was dried (MgSO₄), filtered andconcentrated to dryness to afford about 161 g of Step A compound.

Step B: 3-[2-(tert-Butoxycarbonylamino-methyl)-phenyl]-acrylic acidmethyl ester

To compound of Step A (161.0 g, 561.8 mmol) in DMF (800 mL) was addedmethyl acrylate (58.0 g, 674.2 mmol), TEA (170.5 g, 1685.4 mmol) anddichlorobis(triphenylphosphine) palladium(II) (7.9 g, 11.2 mmol). Themixture was heated at 80° C. for about 32 hours. The mixture was cooled,diluted with 1000 mL of EtOAc and washed with 10% aqueous sodiumbisulfate. The aqueous portion was extracted three times with EtOAc andthe combined organics were dried (Na₂SO₄) and concentrated to dryness.The residue was dissolved in a small amount of DCM and filtered through7 inches of silica gel in a 2 L sintered glass funnel eluting with 25%EtOAc/hexanes. The eluent was concentrated to dryness and recrystallizedfrom EtOAc/hexanes to afford about 116.9 g (71%) of Step B compound.

Step C: To a 0° C. solution of (116.9 g, 401.2 mmol) material from StepB in DCM (800 mL) was added 200 mL of TFA dropwise over 15 minutes.After removing the cooling bath, the mixture was stirred for about 2.5hours and then concentrated to dryness. The residue was dissolved in 500mL of DCM and saturated aqueous sodium bicarbonate is slowly added untilthe mixture was slightly basic. The organic portion was separated andthe aqueous portion is extracted two times with DCM. The combinedorganic portions were dried (Na₂SO₄) and concentrated to dryness. Theresidue was dissolved in 800 mL of DCM and DIPEA (57.0 g, 441.4 mmol)was added. To the mixture was added di-tert-butyl dicarbonate (96.3 g,441.4 mmol) in five portions over 45 minutes and then stirred at r.t.for 16 hours. The mixture was washed with 10% aqueous sodium bisulfate,and the organic portion was separated and the aqueous portion isextracted two times with DCM. The combined organic extracts were dried((Na₂SO₄) and concentrated to dryness. The resulting residue wasdissolved in a small amount of DCM and filtered through 7 inch silicagel in a 2L sintered glass funnel eluting with 25% EtOAc/hexanes. Theeluent was concentrated to dryness and the enantiomers were separated bychiral chromatography. The first eluting isomer was labeled as isomer #1and the second eluting is labeled as isomer #2, which afforded about52.6 g (45%) of the final compound (isomer 2). EIS-MS 292 [M+1].

Preparation 2C 1-Carboxymethyl-1,3-dihydro-isoindole-2-carboxylic acidtert-butyl ester

To 1-methoxycarbonylmethyl-1,3-dihydro-isoindole-2-carboxylic acidtert-butyl ester (52.6 g, 180.5 mmol) in MeOH (500 mL) was added 1 NNaOH (199 mL, 199.0 mmol). The mixture is stirred at r.t. for about 48hours and then concentrated to dryness. The resulting residue wasdissolved in water (300 mL) and extracted with diethyl ether (2×). Theaqueous portion was acidified to pH 2 with 10% aqueous sodium bisulfateand extracted with EtOAc. The combined organic extracts were dried(MgSO₄) and concentrated to dryness to afford about 49.8 g of the finalcompound (99%). EIS-MS 276 [M−1].

Preparation 3C (2-isopropyl-2,3-dihydro-1H-isoindol-1-yl)-acetic acid

Step A: (2,3-dihydro-1H-isoindol-1-yl)-acetic acid methyl ester

To the compound prepared in Preparation C1 (11.75 g., 40.41 mmol) in DCM(50 mL) was added TFA (50 mL) dropwise. After about 2 hours, the mixturewas concentrated to dryness and the resulting residue was partitionedwith saturated aqueous sodium bicarbonate (200 mL) and EtOAc (300 mL).The organic portion was separated and the aqueous layer was extractedwith DCM (4×500 mL). The combined DCM extracts were combined, dried(Na₂SO₄), and concentrated to dryness to afford about 3.97 g (51%).

Step B: (2-isopropyl-2,3-dihydro-1H-isoindol-1-yl)-acetic acid methylester

To the compound obtained from Step A (0.50 g, 2.61 mmol) indichloroethane (46 mL) was added acetone (1.76 mL, 24.01 mmol) andsodium triacetoxyborohydride (2.48 g., 11.74 mmol). After 6 hours, themixture was diluted with 1.0N NaOH (100 mL), and the organic portion wasseparated. The aqueous layer was extracted with DCM (3×100 mL). Thecombined DCM extracts were dried (MgSO₄) and concentrated to dryness toafford about 0.60 g (99%). EIS-MS 235 [M+1].

Step C: To the compound of Step B (0.53 g., 2.30 mmol) in MeOH (5.1 mL)was added 1.0N NaOH (2.53 mL, 2.53 mmol). After two days, the solutionwas concentrated to dryness. The resulting residue was diluted with 1.0NHCl and water was loaded onto a strong cation exchange resin. The resinwas washed with water, THF/water (1:1) and then water. The product wasthen eluted from the resin with pyridine/water (1:9). The eluent wasconcentrated to dryness to afford about 0.43 g (85%) of the finalcompound. EIS-MS 220 [M+1].

Preparation 4C (2-Methyl-2,3-dihydro-1H-isoindol-1-yl)-acetic acid

Step A: (2-Methyl-2,3-dihydro-1H-isoindol-1-yl)-acetic acid methyl ester

The compound from preparation C1 was deprotected with TFA in a mannersimilar to preparation 3C of Step A. To the deprotected compound (0.50g, 2.61 mmol), in dichloroethane (46 mL), was added 37% aqueousformaldehyde solution (1.80 mL, 24.01 mmol) and sodiumtriacetoxyborohydride (2.48 g., 11.74 mmol). After 3 days, the mixturewas diluted with 1.0N NaOH (100 mL). The organic portion was separatedand the aqueous layer was extracted with DCM (3×100 mL). The combinedDCM extracts were dried (Na₂SO₄) and concentrated to dryness. Theresulting residue was purified by flash chromatography (SiO₂, elutingwith 100% EtOAc) affording about 0.43 g (79%) of the alkylatedisoindole. EIS-MS 206 [M+1].

Step B: To the compound of Step A (0.34 g., 1.66 mmol) in MeOH (3.7 ML)was added 1.0N NaOH (1.82 mL, 1.82 mmol). After 2 days, the solution wasconcentrated to dryness. The resulting residue was diluted with 1.0N HCland water was then loaded onto a strong cation exchange resin. The resinwas washed with water, THF/water(1:1) and water, and the product waseluted from the resin with pyridine/water(1:9). The eluent wasconcentrated to dryness to afford about 0.31 g (98%) of the finalcompound. EIS-MS 192 [M+1].

Preparation 5C

The above compound was prepared from Boc-L-Tic-OH as described inPreparation 6C below, except that the Weinreb amide was made by asimilar procedure to that described in Synthesis, 676, 1983.

Preparation 6C

Boc-D-Tic-OH (14.9 g, 53.7 mmol), methoxymethylamine hydrochloride (5.24g, 53.7 mmol), EDC (11.3 g, 59.1 mmol), HOBT (7.98 g, 59.1 mmol), DIEA(9.83 ml, 59.1 mmol) and THF (500 ml) were combined, and the resultingmixture was stirred for about 18 hours at r.t. under nitrogen. Thereaction mixture was concentrated and the residue was taken up in ethylacetate. The resulting mixture was washed with 1M HCl, saturated NaHCO₃and brine, which was then dried via filtration through phase separatorpaper. Removal of solvent gives a residue, which was chromatographed onsilica gel using (1:1 ethylacetate/hexane) to give about 12.3 g ofBoc-D-Tic-NMeOMe (Weinreb amide).

Lithium aluminum hydride (1.0M in THF, 5.1 ml, 5.00 mmol) was slowlyadded to the Weinreb amide prepared above (1.28 g, 4.00 mmol) in THF (35ml) at 0° C. The reaction mixture was stirred at 0° C. for about 15minutes. Aqueous KHSO₄ (970 mg in 20 ml H₂O) was slowly added followedby diethylether. The organic layer was separated and the aqueous layerwas extracted with diethylether. The organic phases were combined andwashed with aqueous 1M HCl, saturated aqueous NaHCO₃ and brine, whichwas then dried over Na₂SO₄. Removal of solvent afforded about 780 mg ofthe final compound. MS: MH+262.

Preparation 7C (2-Butyl-2,3-dihydro-1H-isoindol-1-yl)-acetic acid methylester

The compound from preparation C1 was deprotected with TFA in a mannersimilar to preparation 3C of Step A. To the deprotected compound (0.50g, 2.61 mmol) and butryaldehyde (2.16 mL, 24.01 mmol) in dichloroethane(46 mL) was added sodium triacetoxyborohydride (2.48 g., 11.74 mmol).After reacting about 3 hours, the mixture was diluted with 1.0 N NaOH(100 mL) and partitioned. The aqueous layer was extracted with DCM (3×75mL). The DCM layers were combined, dried over sodium sulfate, filteredand concentrated under reduced pressure to give a brown residue. Theresidue was purified via silica gel chromatography (eluent: ethylacetate/hexanes (1:3). The purified fractions were combined andconcentrated to give the title compound as a brown oil (0.51 g, 77%). MSES 249.2 (M+H)

Preparation 8C (2-Butyl-2,3-dihydro-1H-isoindol-1-yl)-acetic acid

To a solution containing the compound 7C (0.47 g, 1.89 mmol) in methanol(4.2 mL) was added 1.0 N NaOH (2.08 mL, 2.08 mmol). After reacting about2 hours, the solution was concentrated under reduced pressure. Theresidue was diluted with 1.0 N HCl, and water was loaded onto a strongcation exchange resin. The resin was washed with water and THF/water(1:1), and the product was eluted from the resin with pyridine/water(1:9). The pyridine washes were concentrated under reduced pressure, andazeotroped with acetone to give the title compound as brown solids (0.28g.,(64%)) MS ES 234.19 (M+H)

Preparation 9C

Step A: To a solution of N-Boc-4-Fluoro-D-Phe (2.37 g, 8.366 mmol) inmethanol, 3 mL of concentrated sulfuric acid was added. The mixture washeated to reflux overnight and then concentrated in vacuo. MS M+1 198.1

Step B: To an ice cold mixture of 1.65 g (8.367 mmol) of compound fromStep A, 1.353 mL of pyridine and ethyl chloroformate (0.848 mL, 8.869mmol) is added slowly with stirring for about 30 minutes giving whitesolid. The mixture was partitioned between water and ethyl acetate. Theaqueous layer was extracted with EtOAc (2×). The combined organicsolution was dried over MgSO₄, filtered, and concentrated in vacuo togive about 2.17 g of yellow oil (96%). MS M+1 270.1.

Step C: A mixture containing 2.17 g (8.06 mmol) of the compound fromStep B, paraformaldehyde (0.254 g, 8.46 mmol), and 10 mL of 3:1 glacialacetic acid/conc. sulfuric acid was stirred at r.t. for about 48 hours.The mixture was partitioned between water and ethyl acetate. The aqueouslayer was extracted with EtOAc (3×). The combined EtOAc solution wasdried over magnesium sulfate, filtered, and concentrated in vacuo. Thedesired product was purified by column chromatography eluting with 25%EtOAc in Hexane to give about 1.31 g (58%) of colorless oil. MS: M+1282.1

Step D: A solution of 1.31 g (4.656 mmol) of material from Step C in 20mL of 5N HCl was heated to reflux for about 24 hours. The solution wasconcentrated in vacuo. The resulting white solid was washed with etherto afford about 0.87 g (81%). MS M+1 196.1.

Step E: To a solution of 0.87 g (3.755 mmol) of material from Step D in20 ml of 1:1 dioxane/water, di-t-butyl-dicarbonate (0.901 g, 4.131 mmol)and 2.355 mL (16.90 mmol) of TEA were added. The mixture was allowed tostir at r.t. overnight. The mixture was diluted with EtOAc, and theseparated aqueous layer was extracted with EtOAc (3×). The combinedorganic solution was dried over magnesium sulfate, filtered andconcentrated in vacuo to give about 0.64 g (58%)of the final compound.MS M−1 294.1.

Preparation 10C

Step A: By following a procedure of Preparation 28C, Step A and 1.0g(5.58 mmol) of α-methyl-DL-phenylanaline, about 1.4 g of ester wasprepared. MS M+1 194.1

Step B: By following a procedure of Preparation 28C, Step B and 1.08 g(5.59 mmol) of material from Step A, about 1.48 g (100%) of product wasprepared. MS M+1 266.1

Step C: By following a procedure of Preparation 28C, Step C and 1.48 g(5.59 mmol) of material from Step B, about 1.55 g (100%) of product wasprepared. MS M+1 278.1

Step D: By following a procedure of Preparation 28C, Step D and 1.55 g(5.59 mmol) of material from Step C, about 1.33 g of product wasprepared. MS M+1 192.1

Step E: By following a procedure of Preparation 28C, Step E and 1.33 g(5.84 mmol) of material from Step D, about 1.70 g (100%) of the finalcompound was prepared.

MS M+1 292.2

Preparation 11C

Step A: By following a procedure of Preparation 28C, Step A and 2.0g(11.16 mmol) of □-methyl-D-phenylanaline, about 2.15 g of ester wasprepared. MS M+1 194.1

Step B: By following a procedure of Preparation 28C, Step B and 2.15 g(11.16 mmol) of material from Step A, about 1.46 g (49%) of product wasprepared. MS M+1 266.1

Step C: By following a procedure of Preparation 28C, Step C and 1.46 g(5.503 mmol) of material from Step B, about 0.74 g (48%) of product wasprepared. MS M+1 278.1

Step D: By following a procedure of Preparation 28C, Step D and 0.74 g(2.67 mmol) of material from Step C, about 0.54 g (89%) of product wasprepared. MS M+1 192.1

Step E: By following a procedure of Preparation 28C, Step E and 0.54 g(2.37 mmol) of material from Step D, about 0.54 g (78%) of the finalcompound was prepared. MS M+1 292.2

Preparation 12C

Step A: By following a procedure of Preparation 28C, Step A and 0.65 g(1.95 mmol) of N-Boc-4-trifluoromethyl-D-phenylanaline, about 0.48 g ofester was prepared.

MS M+1 248.0

Step B: By following a procedure of Preparation 28C, Step B and 0.48 g(1.95 mmol) of material from Step A, about 0.60 g (96%) of product wasprepared. MS M+1 320.1

Step C: By following a procedure of Preparation 28C, Step C and 0.6 g(1.879 mmol) of material from Step B, about 0.37 g (59%) of product wasprepared. MS M+1 332.1

Step D: By following a procedure of Preparation 28C, Step D and 0.37 g(1.117 mmol) of material from Step C, about 0.11 g (35%) of product wasprepared. MS M+1 246.1

Step E: By following a procedure of Preparation 28C, Step E and 1.11 g(0.391 mmol) of material from Step D, about 0.234 g (>100%) of the finalcompound is prepared. MS M−1 344.1

Preparation 13C Lithium;(2-tert-butoxycarbonyl-1,2,3,4-tetrahydro-isoquinolin-1-yl)-acetate

Step 1: (1,2,3,4-tetrahydro-isoquinolin-1-yl)-acetic acid methyl ester

To a solution 100.4 g (52 mol) of Boc-tetrahydo isoquinoline-1-acetic(100.4 g, 520.0 mmol) in 200 mL methanol was added 400 mL of 2.3 M HClin methanol. The mixture was stirred overnight and concentrated invacuo. The residue was dissolved in ethyl acetate and washed withsaturated sodium bicarbonate, brine, and then dried (Na₂SO₄) andconcentrated in vacuo to afford about 109.5 g (100%) of the titlecompound. EIS-MS: 206 (M+1).

Step 2: 1-methoxycarbonylmethyl-3,4-dihydro-1H-isoguinoline-2-carboxylicacid tert-butyl ester

To a 0° C. solution of material from Step 1 (50.5 g, 240.0 mmol) in 250mL dry THF was added di-tert-butyl dicarbonate (59.3 g, 270.0 mmol) in50 mL dropwise. After siring for about 45 minutes, the mixture wasconcentrated in vacuo. The residue was dissolved in ethyl acetate,washed with saturated sodium bicarbonate and brine, and then dried(Na₂SO₄) and concentrated in vacuo. Chromatography of the residueafforded both enantiomers of the title compound.

EIS-MS: 306 (M+1).

Step 3: To a solution of material from Step 2 (10.2 g, 33.4 mmol) in 220mL of dioxane was added a solution of lithium hydroxide monohydrate(1.67 g, 39.8 mmol) in 110 mL water in portions to maintain atemperature below 30° C. The mixture was stirred for about 16 hours andconcentrated in vacuo to afford about 11.2 g of the final compound.

EIS-MS: 292 (M+1).

Preparation 14C lithium;(2-methyl-1,2,3,4-tetrahydro-isoquinolin-1-yl)-acetate

Step 1: (1,2,3,4-Tetrahydro-isoquinolin-1-yl)-acetic acid methyl ester

The material from Preparation of 13C Step 2 (9.98 g, 32.7 mmol) wasmixed with 500 mL cold 4M HCl/dioxane and stirred at r.t. for about anhour. The mixture was concentrated in vacuo. The residue was dissolvedin ethyl acetate and then washed with saturated sodium bicarbonate andbrine. The organic portion was dried (Na₂SO₄), filtered and concentratedin vacuo to afford about 6.9 g (100%) of the title compound. EIS-MS: 206(M+1).

Step 2: (2-methyl-1,2,3,4-tetrahydro-isoquinolin-1-yl)-acetic acidmethyl ester

To a solution of material from Step 1 (6.71 g, 32.0 mmol) in 175 mL ofdichloroethane was added 37% aqueous formaldehyde (22.6 mL, 300 mmol).After about 10 minute, sodium triacetoxyborohydride (31.2 g, 147.0 mmol)was added in 2 to 3 g portions with some cooling to maintain ambienttemperature. The mixture was stirred for about 16 hours and DCM andwater was added. The mixture was adjusted to pH 9–10 with 5N sodiumhydroxide. The organic layer was separated, washed with brine, and thendried (Na₂SO₄) and concentrated in vacuo. Chromatography (silica gel,5%(2N ammonia in methanol)/DCM) of the residue afforded about 6.9 g(96%) of the title compound. EIS-MS: 220 (M+1).

Step 3: To a solution of material from Step 2 (4.45 g, 18.9 mmol) in 120mL dioxane was added lithium hydroxide monohydrate (1.02 g, 22.7 mmol)in 65 mL water in portions keeping the temperature below 30° C. Afterabout 16 hours, the mixture was concentrated in vacuo to afford about8.12 g of the final compound. EIS-MS: 206 (M+1).

Preparation 15C1,1-Dimethyl-6-methoxy-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acidethyl ester

To a solution of the triflate salt of1,1-dimethyl-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid ethylester (1.5 g, 3.76 mmol, 1.0 eq.) in MeOH (20 mL) and CH₂Cl₂ (2 mL) at0° C. was added a solution of (trimethylsilyl)diazomethane (2.0 M inhexane, 3.7 mL, 2.0 eq.). The resulting mixture was warmed to r.t. andstirred overnight, and then the solution was concentrated. Purificationby flash chromatography (125 g SiO₂ linear gradient, 40 mL/min, 1:1EtOAc/hexane for 33 minutes) afforded about 900 mg of the final compound(96%).

LRMS (electrospray): 250:2 (M+1).

“B Domain” and “C Domain” Combination

Preparation 1BC3-[2-(4-Chloro-phenyl)-1-methoxycarbonyl-ethylcarbamoyl]-3,4-dihydro-1H-isoquinoline-2-carboxylicacid tert-butyl ester (N-Boc-D-Tic-4-Cl-D-phe-OH)

Step 1: The HCl salt of H-D-p-Cl-Phe-OMe (35.8 g, 129 mmol) wasdissolved in water (200 mL). Ethyl acetate (200 mL) was added followedby addition of a saturated sodium bicarbonate solution. The mixture wasstirred for about 5 minutes, and then the organic layer was separated,washed with water (200 mL) and dried over magnesium sulfate.Concentration of the mixture under reduced pressure produces a whitesolid (32.2 g). The solid was then dissolved in methylene chloride (200mL), D-Boc-Tic (35.8 g, 129 mmol) and 4-dimethylaminopyridine (75 mg).The mixture was cooled to 0° C. and JEDC (24.7 g, 129 mmol) was added intwo portions. After stirring for about 20 minutes, the ice bath wasremoved and the solution was allowed to warm to r.t. The solution wasstirred for about 4 hours and then diluted with water (400 mL). Theorganic layer was washed with water (3×), dried over magnesium sulfateand concentrated under reduced pressure to give a clear oil (70 g).Column chromatography (35% ethyl acetate/heptane) afforded about 55.6 gof the intermediate Boc-D-p-Cl-Phe-OMe (85%).

¹H NMR(DMSO) (Two rotomers observed) δ8.26(d, 1H), 8.19(d, 0.5 H),7.24(d, 2H), 7.00–7.19(m, 8H), 4.68(m, 0.5H), 4.20–4.60(m, 4.5H),3.58(s, 3H), 3.51(s, 1.5H), 2.77–3.10(m, 6H), 1.42(s, 3H), 1.21(s, 9H).MS(ES) 473.0(M⁺), 471.1(M⁻).

Step 2: The compound of Step 1 (54.3 g, 114 mmol) was dissolved inmethanol (170 mL). The solution was cooled to 0° C. with an ice bath and1N NaOH (290 mL) is added dropwise. After vigorous stirring for about 20minutes, the mixture was warmed to about 25° C. The solution wasconcentrated under reduced pressure to give yellow oil. The oil wasdissolved in water (200 mL) and the pH is adjusted to about 1. Ethylacetate (200 mL) was added, and the organic layer was separated anddried over magnesium sulfate. Concentration of the organics producedabout 46.3 g of the final compound.

¹H NMRDMSO) (Two rotomers observed) δ7.98(d, 1H), 7.82(d, 0.5 H),6.90–7.41(m, 16H), 4.20–4.70(m, 8.5H), 2.60–3.20(m, 8.5H), 1.32–1.41(m,19H). MS(ES) 459.1 m/z(M⁺), 457.1(M⁻).

Preparation 2BC Boc-L-Tic-4-Cl-D-phe-OH

The above compound was prepared using N-Boc-Tic-OH as described inPreparation 1BC.

¹H NMR(DMSO) (Two rotomers observed) δ7.98(d, 1H), 7.72(d, 0.5 H),6.90–7.41(m, 16H), 4.0–4.70(m, 8.5H), 2.60–3.20(m, 8.5H), 1.32–1.41(m,19H). MS (ES) 459.1 m/z(M⁺), 457.1(M⁻).

Preparation 3BC Lithium;2-[(2-tert-butoxycarbonyl-1,2,3,4-tetrahydro-isoquinolin-3-ylmethyl)-amino]-3-(4-chloro-phenyl)-propionate

Step A: 3-(Methoxy-methyl-carbamoyl)-3,4-dihydro-1Hisoquinoline-2-carboxylic acid tert-butyl ester

To Boc-D-1,2,3,4-tetrahydroisoquinoline carboxylic acid (14.9 g, 53.7mmol), in THF (500 mL), was added N,O-dimethylhydroxylaminehydrochloride (5.24 g, 53.7 mmol), EDC (11.3 g, 59.1 mmol), HOBT (7.98g, 59.1 mmol) and DIPEA (9.83 ml, 56.4 mmol). The mixture was stirredfor about 16 hours at r.t. and under nitrogen, and then concentrated todryness. The resulting residue was taken up in EtOAc, washed with 1MHCl, saturated sodium bicarbonate and brine, and then dried (Na₂SO₄).After concentrating to dryness, the resulting residue was purified byflash chromatography (SiO₂, eluting with 1:1 EtOAc/hexane) to give about12.3 g (71%) of the ester. EIS-MS 321 [M+1]

Step B: 3-Formyl-3,4-dihydro-1H-isoquinoline-2-carboxylic acidtert-butyl ester

To a 0° C. solution of material from Step A (1.28 g, 4.00 mmol) in THF(30 mL) was slowly added 1.0 M LAH (in THF, 5.1 ml, 5.1 mmol). Thereaction mixture was stirred at 0° C. for another 15 minutes. To themixture was slowly added 20 mL of 5% aqueous potassium hydrogensulfateand the mixture extracted with Et₂O (2×). The combined organic portionswere washed with 1M HCl, saturated sodium bicarbonate and brine, andthen dried (Na₂SO₄) and concentrated to dryness affording 0.78 g (75%).

EIS-MS 262 [M+1]

Step C:3-{[2-(4-Chloro-phenyl)-1-methoxycarbonyl-ethylamino]-methyl}-3,4-dihydro-1H-isoguinoline-2-carboxylicacid tert-butyl ester

To a 0° C. solution of 4-Cl-D-Phe-OMe (6.27 g, 25.1 mmol) and sodiumacetate (8.23 g, 100.0 mmol), in 850 ml dry MeOH, was added materialfrom Step B (9.8 g, 37.6 mmol) in 50 ml MeOH. The mixture was stirredfor about 15 minutes and then sodium cyanoborohydride (2.37 g, 37.6mmol) was added. The cooling bath was removed and the reaction stirredfor 16 hours at r.t. The mixture was concentrated to dryness and theresulting residue taken up in water and 1 ml of 1M HCl. The mixture wasextracted with EtOAc, and the organics were washed with saturated sodiumbicarbonate and brine, and then dried (Na₂SO₄) and concentrated todryness. The resulting residue was purified by flash chromatography(SiO₂, eluting with 2:1 hexane/EtOAc) affording about 8.62 g (75%).EIS-MS 459 [M+1]

Step D: To a 12° C. solution of material from Step C (1.11 g, 2.42 mmol)in dioxane (15 ml) was added a solution of lithium hydroxide (0.10 g,2.42 mmol) in water (7.5 mL). The mixture was stirred for about 16 hoursand then concentrated to dryness affording about 1.08 g (100%) of thefinal compound. EIS-MS 445 [M+1].

Preparation 4BC lithium;2-[(2-tert-butoxycarbonyl-1,2,3,4-tetrahydro-isoquinolin-3-ylmethyl)-amino]-3-(4-chloro-phenyl)-propionate

The above compound was Prepared in a manner similar to the preparation3BC above except Boc-L-1,2,3,4-tetrahydroisoquinoline carboxylic acidwas used.

Preparation 5BC Preparation of Lithium2-[(2-tert-butoxycarbonyl-1,2,3,4-tetrahydro-isoquinolin-3-ylmethyl)-methyl-amino]-3-(4-chloro-phenyl)-propionate

Step A: To a solution of3-{[2-(4-Chloro-phenyl)-1-methoxycarbonyl-ethylamino]-methyl}-3,4-dihydro-1H-isoquinoline-2-carboxylicacid tert-butyl ester from preparation 3BC Step C (0.60 gm, 1.31 mmol)in anhydrous methanol, was added sodium acetate (0.54 gm, 6.54 mmol).The solution was brought to pH 5–6 with 3–4 drops of glacial aceticacid. Aqueous formaldehyde (37% by wt., 0.49 mL) was added. The solutionwas put under a nitrogen atmosphere and cooled to 0° C. After about 15minutes, sodium cyanoborohydride (0.25 gm, 3.92 mmol) was added andrinsed into the reaction with anhydrous methanol (5 mL). The mixture wasstirred at r.t. overnight, and then concentrated in vacuo andreconstituted in aqueous sodium bicarbonate and ethyl acetate. Afterseparation of phases, the aqueous phase was extracted with ethyl acetate(2×), and all organics were combined, dried (magnesium sulfate),filtered, and concentrated to an opaque white oil (0.64 gm).Chromatography (0 to 20% ethyl acetate in hexane) gave about 0.6 g ofmethylated product as a clear oil (97%). MS (m/z, ES+): 473.2.

Step B: A solution of LiOH.H₂O (0.05 gm, 1.27 mmol) in distilled water(4 mL) was added to a solution of the material from Step A in1,4-dioxane (8 mL), and the reaction was cooled slightly in an ice waterbath. The mixture was stirred under a nitrogen atmosphere at r.t.overnight. An additional 1.5 eq. of LiOH.H₂O (0.08 gm) were added as anaqueous solution (4 mL), and the mixture was stirred at r.t. over theweekend. The mixture was concentrated, and then combined with THF andconcentrated (3×) to help dry the material. The resulting foam was driedat r.t. overnight in a vacuum oven to give about 0.67 g of finalcompound as a white foam (114%). MS (m/z, ES+): 459.2

Preparation 6BC lithium2-[(2-tert-butoxycarbonyl-1,2,3,4-tetrahydro-isoquinolin-3-ylmethyl)-(2-methoxy-ethyl)-amino]-3-(4-chloro-phenyl)-propionate

Step A: To a solution of methoxyacetaldehyde (0.15 gm, 2.03 mmol),3-{[2-(4-Chloro-phenyl)-1-methoxycarbonyl-ethylamino]-methyl}-3,4-dihydro-1H-isoquinoline-2-carboxylicacid tert-butyl ester from preparation 3BC Step C (0.31 gm, 0.68 mmol)in acetonitrile was added sodium triacetoxyborohydride (0.72 gm, 3.38mmol). After stirring overnight under a nitrogen atmosphere at r.t.,additional acetaldehyde (0.25 gm) dissolved in acetonitrile and sodiumtriacetoxyborohydride (0.21 gm) was added, and the mixture was stirredfor about 8.5 hours. The mixture was quenched at r.t. with 5N NaOH (5mL). The aqueous phase was separated from the organic and extracted withethyl acetate (4×). The combined organics were washed with a brinesolution, and then dried, filtered and concentrated. Chromatography(gradient of ethyl acetate in hexane, 0 to 12%) gives about 0.23 g of3-{[[2-(4-Chloro-phenyl)-1-methoxycarbonyl-ethyl]-(2-methoxy-ethyl)-amino]-methyl}-3,4-dihydro-1H-isoquinoline-2-carboxylicacid tert-butyl ester as a yellow oil (70%). MS (m/z, ES+): 517.2.

Step B: To a solution of the material from Step A in 1,4-dioxane wasadded a solution of lithium hydroxide monohydrate (0.05 gm, 1.11 mmol)in distilled water (2 mL). The mixture was stirred overnight at r.t. andthen concentrated to a white residue. Addition of THF and concentration(3×) gives the lithium carboxylate as a foam. The foam was driedovernight under vacuum to afford about 0.25 g of crude solids (109%). MS(m/z, ES+): 503.3

Preparation 7BC1-{[1-Carboxy-2-(4-chloro-phenyl)-ethylcarbamoyl]-methyl}-1,3-dihydro-isoindole-2-carboxylicacid tert-butyl ester

Step A: To a suspension of 4-Cl-D-Phe-OMe hydrochloride (40.4 g, 161.5mmol) in DCM (250 mL) was added saturated aqueous sodium bicarbonate(250 mL), and the mixture was stirred at r.t. for about 1 hour. Theorganic portion was separated and the aqueous portion was extracted withDCM (2×). The combined organic portions were dried (Na₂SO₄) andconcentrated to dryness. To the free amine, in DCM (400 mL) at 0° C.,was added 1-Carboxymethyl-1,3-dihydro-isoindole-2-carboxylic acidtert-butyl ester from preparation 2C (isomer 2, 44.8 g, 161.5 mmol), EDC(31.0 g, 161.5 mmol) and 4-DMAP (2.0 g, 16.1 mmol). The mixture wasstirred at 0° C. for about 30 minutes whereupon the cooling bath wasremoved and the mixture was stirred for another 5 hours at r.t. Themixture was then washed with saturated aqueous sodium bicarbonate (200mL) and 10% aqueous sodium bisulfate (200 mL), and then dried (Na₂SO₄)and concentrated to dryness to afford about 76.4 g (100%) of the ester.EIS-MS 471 [M−1]

Step B: To the ester from Step A (76.4 g, 161.5 mmol) in MeOH (760 mL)was added 1 N NaOH (242.0 mL, 242.0 mmol), and the mixture was heated at50° C. for 4 hours and then stirred for another 16 hours at r.t. Afterconcentrating to dryness, the resulting residue was taken up in 500 mLof water and washed with diethyl ether (2×). The aqueous portion wasacidified to pH 2 with 10% aqueous sodium bisulfate and extracted withEtOAc (4×200 mL). The combined organic extracts were dried (MgSO₄) andconcentrated to dryness. The resulting solid was suspended in hexanes,filtered, and dried to afford about 67.7 g (91%) of the final compound.EIS-MS: 457 [M−1]

Preparation 8BC3-(4-Chloro-phenyl)-2-[(1,1-dimethyl-1,2,3,4-tetrahydro-isoquinoline-3-carbonyl)-amino]-propionicacid methyl ester

To a solution of 1,1-dimethyl Tic (240 mg, 1.17 mmol), 4-Cl-D-Phe-OMe(322 mg, 1.28 mmol), HOBT (197 mg, 1.46 mmol), and DIPEA (0.81 mL, 44.68mmol) in DCM/DMF (1:1) was added EDC (280 mg, 1.46 mmol). The resultingmixture was stirred at r.t. overnight. The mixture was then diluted withEtOAc (100 mL), washed with saturated aqueous NaHCO₃ and brine, and thendried (Na₂SO₄) and concentrated to dryness. Purification and separationof diastereomers by flash chromatography (35 g SiO₂, linear gradient, 40mL/min 10–50% EtOAc/hexane for 25 minutes and 50% EtOAc/hexane for 7minutes) afforded the final compound. LRMS (ESI+): 401.1 (M+H)

Preparation 9BC3-(4-Chloro-phenyl)-2-[(1,1-dimethyl-1,2,3,4-tetrahydro-isoquinoline-3-carbonyl)-amino]-propionicacid

To the compound from preparation 8BC (5.95 g, 14.88 mmol) in a 1:1mixture of THF/H₂O (50 mL) was added lithium hydroxide hydrate (0.75 g,17.87 mmol). The mixture was stirred at r.t. for about 18 hours. Themixture was then concentrated to dryness. The resulting residue wasdissolved in water (50 mL), made acidic with 1N HCl (25 mL) and washedwith Et₂O (100 mL). The aqueous layer was evaporated to dryness toafford about 6.18 g of the final compound (98%). LRMS(EIS+): 387 [M+1]

Preparation 10BC1-{[1-Carboxy-2-(4-methoxy-phenyl)-ethylcarbamoyl]-methyl}-1,3-dihydro-isoindole-2-carboxylicacid tert-butyl ester

Step 1: To a solution of p-methoxy-D-Phe-OMe (1.72 g, 8.23 mmol)dissolved in THF (45 mL) and1-carboxymethyl-1,3-dihydro-isoindole-2-carboxylic acid tert-butyl ester(2.51 g, 9.05 mmol) was added HOBT (1.22 g, 9.05 mmol), EDC (1.73 g,9.05 mmol) and DIPEA (1.6 mL, 9.05 mmol). The reaction was stirredovernight at r.t. and then concentrated. The mixture was washed with 1MHCl, dilute NaHCO₃ and brine, and then dried with sodium sulfate. Themixture was chromatographed on silica gel eluting with 3% 2M NH₃ inMeOH/CH₂Cl₂ giving about 2.58 g as white solids. Mass MH⁺ 469

Step 2: The white solid from Step 1 (2.58 g, 5.5 mmol) was dissolved indioxane (37 mL) and lithium hydroxide hydrate (0.35 g, 8.3 mmol) in H₂O(19 mL) was added. The mixture was stirred for about 2.5 hours at r.t.and then concentrated. Ethyl acetate was added and the mixture waswashed with 1M HCl and brine, and then concentrated to afford about 2.56g of the final free acid. LRMS(ESI+): 455 (M+1)

Preparation 11BC1-[1-Carboxy-2-(4-chloro-phenyl)-ethylcarbamoyl]-1,3-dihydro-isoindole-2-carboxylicacid tert-butyl ester

Step 1: About 2.0 g (7.60 mmol) of (R,S)-Boc-1,3-dihydro-2H isoindolecarboxylic acid was dissolved in 100 ml THF and about 2.28 g (9.12 mmol)of 4-Cl-D-phe-methylester HCl, 1.25 g (9.12 mmol) of HOBT, 1.75 g (9.12mmol) of EDC, and 1.6 ml (9.12 mmol) of DIEA were added. The mixture wasstirred overnight at r.t., concentrated to dryness, washed with 1M HCl,dilute NaHCO₃ and brine, and then dried over sodium sulfate. Thematerial was chromatographed on silica gel by eluting with ethylacetate/hexane 1:2 to give about 1.05 g of isomer 1 and about 0.82 g ofisomer 2, and about 1.61 g mixture of isomers 1 and 2. Mass MH⁺ 459

Step 2: About 0.82 g (1.79 mmol) of the isomer 2 obtained in Step 1 wasdissolved in 11 ml of dioxane and 0.11 g (2.68 mmole) of LiOH-hydrate in5.5 ml of H₂O was added. The mixture was stirred for about 4 hours atr.t. and then concentrated to dryness. Ethyl acetate was added, and thesolution was washed with 1M HCl and brine, and then concentrated todryness affording about 0.75 g of the free acid. Mass: 445 (MH⁺)

EXAMPLE Example 1 Coupling Procedure 16-Methoxy-1,1-dimethyl-1,2,3,4-tetrahydro-isoguinoline-3-carboxylic acid(1-(4-chloro-benzyl)-2-{4-[2-(2-morpholin-4-yl-ethoxy)-phenyl]-piperazin-1-yl}-2-oxo-ethyl)-amidedichloride

Step 1: To a 3000 mL flask containing4-[2-(2-Piperazin-1-yl-phenoxy)-ethyl]-morpholine (25.3 g, 0.0868 mol),Boc-D-p-Cl-Phe (28.6 g, 0.0954 mol), HOBT (13.5 g, 0.10 mol), iPr₂NEt(30.2 mL, 0.173 mol), 800 mL of CH₂Cl₂, and 200 mL of DMF was added EDC(19.1 g, 0.10 mol). After stirring overnight, the solution wasconcentrated to remove the CH₂Cl₂ and divided into two equal portionswhich were each diluted with 1000 mL of EtOAc. The organic solutionswere washed with saturated sodium bicarbonate, water (2×) and brine, andthen dried (Na₂SO₄), filtered and concentrated. Half of the material wassubjected to silica gel chromatography (0 to 5% MeOH/CH₂Cl₂) to afford awhite solid. The other half was dissolved in Et₂O and precipitated byadding 1 M HCl in Et₂O. The precipitate was washed with Et₂O,transferred to a flask as a slurry in Et₂O, and concentrated to afford awhite solid. The Boc protected product purified by flash chromatographywas deprotected as described in Step 2 below. The Boc protected productpurified by precipitation was deprotected in a similar manner. Thepurity of the two batches of material were identical by HPLC (>99%).Combined yield: 38.4 g, 0.070 mol, 81%.

Step 2: To a solution of(1-(4-chloro-benzyl)-2-{4-[2-(2-morpholin-4-yl-ethoxy)-phenyl]-piperazine-1-yl}-2-oxo-ethyl)-carbamicacid tert-butyl ester (23.57 g, 41.1 mmol) in MeOH (225 mL) was added1.0 M HCl in Et₂O. The mixture is stirred at r.t. overnight. The solidwas filtered, washed with Et₂O and dried under vacuum overnight toafford2-amino-3-(4-chloro-phenyl)-1-{4-[2-(2-morpholin-4-yl-ethoxy)-phenyl]-piperazine-1-yl}-propan-1-one(19.6 g, 36 mmol, 88%).

Step 3: To a solution of compound from Step 2 (2.0 g, 3.69 mmol, 1.1eq.), 1,1-dimethyl-6-methoxy-1,2,3,4-tetrahydroisoquinoline-3-carboxylicacid (810 mg, 3.36 mmol, 1.0 eq.), HOBT (567 mg, 4.2 mmol, 1.25 eq.),DIPEA (2.35 mL, 13.44 mmol, 4.0 eq.), CH₂Cl₂ (20 mL), and DMF (20 mL)was added 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride(805 mg, 4.2 mmol, 1.25 eq.). The mixture was stirred at r.t. overnightand diluted with ethyl acetate. The mixture was washed with saturatedaqueous sodium bicarbonate and brine, and then dried over Na₂SO₄,filtered, and concentrated. Purification by flash chromatography (125 gSiO₂, linear gradient, 40 mL/minute, 0 to 10% MeOH/CH₂Cl₂ over 20minutes and 10% MeOH/CH₂Cl₂ for 13 minutes) afforded the final compound(1.68 g, 2.4 mmol, 73%) as a mixture of diastereomers. The finalcompound was converted to chloride salt by adding 1.0 M HCl in Et₂O. Thetwo diastereomers were separated by reversed-phase chromatography.

Isomer-1 (667463): HRMS (electrospray) calcd for C₃₈H₄₈N₅O₅Na Cl:712.3242. Found: 712.3253.

Isomer-2: HRMS (electrospray) calcd for C₃₈H₄₈N₅O₅ Na Cl: 712.3242.Found: 712.3278.

Example 2 Coupling Procedure 2N-[1-(4-Chloro-benzyl)-2-oxo-2-(4-{2-[1-(2,2,2-trifluoro-ethyl)-piperidin-3-yloxy]-phenyl}-piperazin-1-yl)-ethyl]-2-(2,3-dihydro-1H-isoindol-1-yl)-acetamidetristrifluoroacetic Acid

Step 1: To a solution of 208 mg (0.455 mmol, 1.2 eq.)1-{[1-carboxy-2-(4-chloro-phenyl)-ethylcarbamoyl]-methyl}1–1,3-dihydro-isoindole-2-carboxylicacid tert-butyl ester, 0.13 mL, (0.758 mmol, 2.0 eq.) of DIPEA, and 173mg (0.455 mmol, 1.2 eq.) of HATU in 8 mL of DCM and 1 mL of DMF wasadded a solution of 130 mg (0.379 mmol, 1.0 eq.) of1-{2-[1-(2,2,2-trifluoro-ethyl)-piperidin-3-yloxy]-phenyl}-piperazine in2 mL of DCM. The solution was stirred at r.t. for about 3 hours and thenconcentrated to an oil. Ethyl acetate was added and the solution waswashed with saturated sodium bicarbonate solution, water (4×) and brine,and then dried with sodium sulfate, filtered, and concentrated to anoil. Purification by flash chromatography eluting with 3% NH₃ (2M) inmethanol/DCM afforded about 120 mg of oily residue. MS: (M+1) 784

Step 2: The residue was dissolved in 1 mL of DCM, cooled with an icebath, and 2 mL of cooled TFA/DCM (1/1) was added. The mixture wasstirred for about 1 hour with an ice bath in place. The mixture wasconcentrated and purified as the TFA salt via reverse phase HPLC givingabout 10.6 mg of the final compound (7%). HRMS (electrospray) calculatedfor C₃₆H₄₁ClF₃N₅O₃3C₂HF₃O₂: 684.2928. Found: 684.2932.

Example 3 Coupling Procedure 3 [3R,3(1R)]-1,2,3,4-Tetrahydro-isoquinoline-3-carboxylic acid(1-(4-chloro-benzyl)-2-{4-[2-(1-methyl-piperidin-3-yloxy)-phenyl]-piperazin-1-yl}-2-oxo-ethyl)-amidetristrifluoroacetate

Step 1: 1-[2-(1-Methyl-piperidin-3-yloxy)-phenyl]-piperazine(0.20 g,0.73 mmol) was taken up in methylene chloride (30 mL) and treated withHOBT (0.10 g, 0.73 mmol),3-[1-carboxy-2-(4-chloro-phenyl)-ethylcarbamoyl]-3,4-dihydro-1H-isoquinoline-2-carboxylicacid tert-butyl ester (0.37 g, 0.80 mmol), and1,3-dicyclohexylcarbodiimide (0.17 g, 0.80 mmol). The mixture wasstirred at r.t. for about 30 minutes. The precipitate was filtered off,and the solution was diluted with ethyl acetate (400 mL) and washed withwater (60 mL), saturated aqueous sodium bicarbonate (60 mL) and brine(60 mL), and then dried over anhydrous sodium sulfate. The solvent wasconcentrated under reduced pressure, and the resulting residue waspurified via silica gel chromatography (5% methanol in ethyl acetate).

Step 2: The resulting product was dissolved in methylene chloride (5mL), and TFA(5 mL) was added. The mixture was stirred for about 2 hours.The solvent was concentrated under reduced pressure and the residue waspurified via preparatory HPLC to afford the final compound (0.80 g, 10%)as a white solid.

¹H NMR (CD₃OD) δ 7.24–7.47 (m, 8H), 6.88–7.18 (m, 4H), 5.17–5.30 (m,1H), 4.404–4.51 (s, 2H), 4.17–4.30 (m, 1H), 3.57–3.83 (m, 5H), 3.38–3.55(m, 3H), 2.95–3.18 (m, 7H), 2.92 (s, 3H), 1.63–1.85 (m, 6H). MS (ESI):m/z=616 [C₃₅H₄₂ClN₅O₃+H]⁺

Example 4 Coupling Procedure 4N-(1-(4D-chloro-benzyl)-2-oxo-2-{4-[2-R-(piperidin-3-yloxy)-phenyl]-piperazin-1-yl}-ethyl)-2-(1,2,3,4-tetrahydro-isoquinolin-1-yl)-acetamidetrihydrochloride

Step 1: 1-[2-(1-Boc-R-piperidin-3-yloxy)-phenyl piperazine (3.91 g,10.82 mmol, 1 eq.), HOBT (1.46 g, 10.82 mmol, 1 eq.), DIPEA (5.67 mL,10.82 mmol, 1 eq.) and FMOC D-Cl Phe (4.57 g, 10.82 mmol) were mixedtogether in DCM (180 mL) and DMP (20 mL). EDC (2.08 g, 10.82 mmol, 1eq.) was added, and the mixture was stirred at r.t. for about 16 hours.The mixture was concentrated to a yellow foam. Chromatography on silicagel gave an off white foam (6.68 g, 80%).

MS found: 765.2 M+1

Step 2: The material from Step 1 (6.62 g, 8.65 mmol) was dissolved inTBF (300 mL), and TBAF (17.3 mL of 1 M solution in THF, 17.3 mmol) wasadded at 0° C. while stirring. The mixture was stirred for about an hourand concentrated to a thick oil. The material was dissolved in ethylacetate (200 mL) and washed with water (2×200 mL). The organic fractionwas dried over sodium sulfate, filtered and concentrated. Silica gelchromatography gave2-amino-3D-(4-chloro-phenyl)-1-{4-[2R-(piperidin-3-yloxy)-phenyl]-piperazine-1-yl}-propan-1-one(4.5 g, 97%) as a yellow foam.

MS found 543.2 M+1

Step 3: To a solution of the compound from Step 2 (130 mg, 0.21 mmol,1.0 eq.), lithium salt of1-carbomethyl-3,4-dihydro-1H-isoquinoline-2-carboxylic acid tert-butylester (75 mg, 0.25 mmol, 1.2 eq.), HOBT (38 mg, 0.25 mmol, 1.2 eq.),DIPEA (0.11 mL, 0.63 mmol, 3.0 eq.), CH₂Cl₂ (4 mL) and DMF (2 mL) wasadded EDC (48 mg, 0.25 mmol, 1.2 eq.). The mixture was stirred at r.t.overnight. The mixture was diluted with ethyl acetate and washed withsaturated aqueous sodium bicarbonate and brine, and then dried overNa₂SO₄, filtered and concentrated. Purification by flash chromatography(10 g SiO₂, linear gradient, 40 mL/minute, 0 to 10% MeOH/CH₂Cl₂ over 25minutes and 10% MeOH/CH₂Cl₂ for 7 minutes) afforded boc-protected titlecompound (168 mg, 0.2 mmol, 98%).

Step 4: To a solution of boc-protected title compound (155 mg, 0.19mmol) in CH₂Cl₂ (2 mL) was added TFA (2 mL) and DMS (0.25 mL). Themixture was stirred at r.t. for about 2 hours. The mixture wasconcentrated and purified using SCX (10 g) ion-exchange chromatographyto afford final compound (121 mg, 0.16 mmol, 88%), which was convertedto chloride salt by adding 1.0 M HCl in Et₂O.

HRMS (electrospray) calculated for C₃₅H₄₃N₅O₃Cl: 616.3054. Found:616.3073.

Example 5 Coupling Procedure 53-D-(4-chloro-phenyl)-1-{4-[5-trifluoromethyl-2-S-(pyrrolidin-3-yloxy)-phenyl]-piperazin-1-yl}2-D-[(1,2,3,4-tetrahydro-isoquinoline-3-ylmethyl)-amino]-propan-1-one3HCl Salt

Step 1: To a solution of lithium;2-[(2-tert-butoxycarbonyl-1,2,3,4-tetrahydro-isoquinolin-3-ylmethyl)-amino]-3-(4-chloro-phenyl)-propionate(359mg, 0.79 mmol, 1.2 eq),3-(2-piperazin-1-yl-trifluoromethyl-phenoxy)-S-pyrrolidine-1-carboxylicacid tert-butyl ester (275 mg, 0.66 mmol, 1.0 eq.), DIPEA (0.576 mL, 3.3mmol, 5 eq.), CH₂Cl₂ (18 mL), HOBT (107 mg, 0.79 mmol, 1.2 eq.), and DMF(2 mL), was added EDC (151 mg, 0.79 mmol, 1.2 eq.). The solution wasstirred at r.t. for about 16 hours and then was concentrated to an oil.

Step 2: The residue was taken up in DCM/TFA 1/1 (10 mL) and stirred atr.t. for about 16 hours. The mixture was concentrated and free based viaSCX ion exchange chromatography. The product containing fractions wasconcentrated to give an oily residue. Chromatography on silica gelfollowed by addition of excess HC1 in diethyl ether gave the finalcompound (395 mg, 84%) as an off white solid. MS found 643.3

Example 6 Coupling Procedure 61,2,3,4-tetrahydro-isoquinoline-3-D-carboxylic acid(1-D-(4-chloro-benzyl)2-oxo-2-S-{4-[2-NH-(piperidin-3-yloxy)-phenyl]-piperadin-1-yl}-ethyl)-amide2HCl Salt

Step 1: To a solution of3-[1-carboxy-2-(4-chloro-phenyl)-ethylcarbamoyl]-3,4-dihydro-1H-isoquinoline-2-carboxylicacid tert-butyl ester (693 mg, 1.51 mmol, 1.1 eq.),2,2,2-trifluoro-1-[3-(2-S-piperidin-4-y1-phenoxy)-piperidin-1-yl]-ethanone(489 mg, 1.37 mmol, 1.0 eq.), DIPEA (0.79 mL, 4.52 mmol, 3.3 eq.) andCH₂Cl₂ (10 mL) was added HATU (574 mg, 1.51 mmol, 1.1 eq.). The solutionwas stirred at r.t. for about 16 hours and concentrated to an oil.Purification by flash chromatography afforded an oily residue.

Step 2: The residue was taken up in 7 N NH₃/MeOH (15 mL) and stirred atr.t. for about 12 hours. The mixture was concentrated, and the residuewas dissolved in TFA/DCM (1/1) 15 mL) and stirred at r.t. for about 12hours. The residue was freebased via SCX ion exchange chromatography.The product containing fractions was concentrated to give an oilyresidue. Chromatography on silica gel followed by addition of excess HClin diethyl ether gave the final compound (358 mg, 53%) as an off whitesolid.

MS found 601.1

Examples 7–48

The Examples 7–48 are prepared from an appropriate A domain piperazineby following a substantially similar coupling procedure as described inProcedures 1–6 (Examples 1–6).

Example Z Coupling Procedure MS (ESI) 7

2 604.3 (M + H) 8

2 604.3 (M + H) 9

2 646.3 (M + H) 10

2 604.3 (M + H) 11

2 616.3  M + H) 12

5 602.3 (M + H) 13

5 602.3 (M + H) 14

1 604.3 (M + H) 15

2 632.0 (M + H) 16

3 616.0 (M + H) 17

2 680.0 (M + H) 18

2 630.0 (M + H) 19

2 616.0 (M + H) 20

2 644.0 (M + H) 21

2 655.0 (M + H) 22

2 668.0 (M + H) 23

2 709.0 (M + H) 24

2 652.0 (M + H) 25

2 632.0 (M + H) 26

2 588.0 (M + H) 27

2 706.0 (M + H) 28

2 672.0 (M + H) 29

2 742.0 (M + H) 30

2 708.0 (M + H) 31

2 630.0 (M + H) 32

2 630.0 (M + H) 33

2 694.0 (M + H) 34

2 658.0 (M + H) 35

2 616.0 (M + H) 36

5 602.0 (M + H) 37

2 644.0 (M + H) 38

2 590.3 (M + H) 39

2 602.0 (M + H) 40

2 617.0 (M + H) 41

2 644.0 (M + H) 42

2 630.3 (M + H) 43

2 602.3 (M + H) 44

2 602.3 (M + H) 45

5 588.3 (M + H) 46

5 588.3 (M + H) 47

2 616.3 (M + H) 48

2 646.3 (M + H)

Examples 49–50

The compounds of Examples 49–50 are prepared from an appropriate Adomain piperazine by following a substantially similar couplingprocedure as described in Procedures 1–6 (Examples 1–6).

Coupling Example Z Procedure MS (M + H) 49

1 616.3 50

4 602.3

Examples 51–57

Examples 51–57 are prepared from an appropriate A domain piperazine byfollowing a substantially similar coupling procedure as described inProcedures 1–6 (Examples 1–6).

Exam- Coupling MS ple Z Procedure (M + H) 51

5 630.3 52

2 644.3 53

2 644.3 54

5 630.3 55

5 682.3 56

2 643.3 57

5 630.3

Examples 58–66

Examples 58–66 are prepared from an appropriate A domain piperazine byfollowing by following a substantially similar coupling procedure asdescribed in Procedure 5 (Example 5).

Exam- Coupling ple Z Procedure MS (ESI) 58

5 588.3(M + H) 59

5 606.3(M + H) 60

5 606.3(M + H) 61

5 642.3(M + H) 62

5 606.3(M + H) 63

5 606.3(M + H) 64

5 588.3(M + H) 65

5 616.3(M + H) 66

5 608.3(M + H)

Examples 67–85

Examples 67–85 are prepared from an appropriate A domain piperazine byfollowing a substantially similar coupling procedure as described inProcedures 1–6 (Examples 1–6).

Example Z Coupling Procedure MS (ESI) 67

1 682.3 (M + H) 68

1 633.0 (M + H) 69

1 696.3 (M + H) 70

5 602.4 (M + H) 71

2 632.2 (M + H) 72

2 644.3 (M + H) 73

2 644.3 (M + H) 74

2 308.7 (M + 2H)/2 75

1 638.3 (M + H) 76

4 602.3 (M + H) 77

2 658.5 (M + H) 78

2 672.4 (M + H) 79

2 660.3 (M + H) 80

2 674.3 (M + H) 81

2 688.3 (M + H) 82

2 680.3 (M + H) 83

2 694.3 (M + H) 84

2 708.3 (M + H) 85

2 684.3 (M + H)

Examples 86–88

Examples 86–88 are prepared from an appropriate A domain piperazine byfollowing a substantially similar coupling procedure as described inProcedures 1–6 (Examples 1–6).

Example Z Coupling Procedure MS (ESI) 86

1 630.3 (M + H) 87

4 616.3 (M + H) 88

1 646.4 (M + H)

Examples 89–90

Examples 89–90 are prepared from an appropriate A domain piperazine byfollowing a substantially similar procedure as described in CouplingProcedure 1 (Example 1).

Ex- am- Coupling ple Z Procedure MS (ESI) 89

1 660 (M + H) 90

1 660 (M + H)

Examples 91–98

Examples 91–98 are prepared from an appropriate A domain piperazine by asubstantially similar procedure as described in Coupling Procedures 1–6(Examples 1–6).

Example Z Coupling Procedure MS (ESI) 91

1 646.4 (M + H) 92

1 646.4 (M + H) 93

1 652.3 (M + H) 94

1 630.3 (M + H) 95

4 616.3 (M + H) 96

4 616.3 (M + H) 97

5 616.2 98

5 616.2

Examples 99

Example 99 is prepared from an appropriately substituted A domainpiperazine by following a substantially similar coupling procedure asdescribed in Coupling Procedure 5 (Example 5).

Found HRMS (electrospray): 588.2763 (M+H)

Examples 100–101

Examples 101–102 are prepared from an appropriately substituted A domainpiperazine by following a substantially similar coupling procedure asdescribed in Coupling Procedures 1–6 (Examples 1–6).

Found Coupling HRMS Example Z Procedure (electrospray) 100

5 598.3412(M + H) 101

2 612.3570(M + H)

Example 102

Example 103 is prepared from an appropriately substituted A domainpiperazine by following a substantially similar coupling procedure asdescribed in Coupling Procedure 5 (Example 5).

Found HRMS (electrospray): 616.3037 (M+H)

Example 103

Example 103 is prepared from an appropriately substituted A domainpiperazine by following substantially similar coupling procedure asdescribed in Coupling Procedure 1 (Example 1).

Found HRMS (electrospray): 588.3253 (M+H)

Examples 104–107

Examples 104–107 are prepared from an appropriately substituted A domainpiperidine by following a substantially similar coupling procedure asdescribed in Coupling Procedures 1–6 (Examples 1–6).

Example Z Coupling Procedure MS (ESI) 104

6 601.3 (M + H) 105

2 615.3 (M + H) 106

2 631.3 (M + H) 107

6 587.3 (M + H)

Examples 108–109

Examples 108–109 are prepared from an appropriately substituted A domainpiperidine by following a substantially similar coupling procedure asdescribed in Coupling Procedures 1–6(Examples 1–6).

Ex- am- Coupling ple Z Procedure MS (ESI) 108

2 631.3 (M + H) 109

6 601.2 (M + H)

Examples 110–111

Examples 110–111 are prepared from an appropriately substituted A domainpiperidine by following a substantially similar coupling procedure asdescribed in Coupling Procedures 1–6 (Examples 1–6).

Coupling Example Z Procedure MS (ESI) 110

6 629.3 (M + H) 111

2 643.2 (M + H)

Examples 112–115

The compounds of Examples 112–115 are prepared from an appropriatelysubstituted A domain piperazine by following a substantially similarcoupling procedure as described in Coupling Procedures 1–5.

Example Z Coupling Procedure MS (ESI) 112

4 598.3 (M + H) 113

1 628.6 (M + H) 114

2 628.1 (M + H)Preparation of Novel C-Domain PiecesHeck Coupling

Preparation PP1

Synthesis of Compound (2a) by a Heck Coupling of 2-bromobenzaldehyde(1a) with methyl acrylate (Pd(OAc)₂/PPh₃ as the catalyst): A mixture of2-bromobenzaldehyde (1a) (24.5 g, 132 mmol), methyl acrylate (17.9 mL,199 mmol), Pd(OAc)₂ (590 mg, 2.65 mmol, 2 mol %), PPh₃ (1.39 g, 5.30mmol, 4 mol %) and Et₃N (46 mL, 331 mmol) was stirred at 80° C. for 15h. Large amount of yellow solid was formed after the reaction was done.The mixture was cooled to rt, concentrated, and mixed with H₂O (200 mL).The organic solid was collected by filtration, and then applied to aplug of silica gel (25 g) (EtOAc/hexane 1:1) to give a dark yellowsolid. The solid was purified by crystallization (100 mL EtOAc bottomlayer, 120 mL hexane top layer) to provide 17.57 g (70%) (100% pure byNMR) of the first crop and 5.23 g (21%) (95% by NMR) of the second cropof 2a.

Preparation PP2

Synthesis of Compound (2a) by a Heck Coupling of 2-bromobenzaldehyde(1a) with Methyl Acrylate (R═H) (Pd(OAc)₂/P(O-Toyl)₃ as the catalyst):The compound 1a (9.998 g, 54.04 mmol) was dissolved in toluene (20 mL)at r.t. Methylacrylate (5.996 g, 69.65 mmol, 1.29 eq.), NEt₃ (15 mL),Pd(OAc)₂ and P(O-Tolyl)₃ were successively added and the mixture wasstirred under reflux. After 2 hours, the reaction mixture was allowed tocool to RT. Then the precipitated yellow catalyst was removed byfiltration. The catalyst was rinsed with toluene (2×10 mL) and thefiltrates were concentrated to dryness under reduced pressure. Theresidual oil was dried under vacuum over the weekend to give a crudesolid (11.449 g). The solid was taken-up with isopropanol (25 mL) andstirred overnight at RT. Then, the precipitate was filtered and rinsedwith isopropanol (5 mL). The wet cake (8.240 g) was dried overnight atRT affording the highly pure 2-carboxaldehyde-methyl-cinnamate with 74%yield (7.627 g, 40.1 mmol).

Preparation PP3

Heck Coupling of 1b and methyl acrylate to form 2b (R=5-OMe): A mixtureof 2-bromo-5-methoxybenzaldehyde (1b) (4.5 g, 20.9 mmol, Aldrich),methyl acrylate (2.7 g, 1.5 eq, 2.83 mL), Et₃N (7.4 g, 3.5 eq, 10.2 mL),Pd(OAc)₂ (93 mg, 0.02 eq), and P(O-Tol)₃ was stirred and heated to 80°C. over 2–3 days. The reaction mixture was cooled to r.t., partitionedbetween EtOAc (50 mL) and brine (50 mL). The aqueous was extracted withEtOAc (2×50 mL). The combined organic was washed with brine (1×50 mL),dried over MgSO₄, filtered, concentrated to yield a yellow brown oil(5.01 g, 109%). This crude oil was purified in a hot solvent Hex/EtOAc(80 mL/15 mL) to yield 2b as a pale yellow solid (3.5 g, 76%).

Preparation PP4

Heck Coupling of 1c and Methyl Acrylate to Form 2c (R=4,5-OMe): To asolution of 1c (906 mg, 3.70 mmol) in toluene (2 mL) was added Pd(OAc)₂(17 mg, 0.074 mmol, 2 mol %), P(O-Tolyl)₃ (45 mg, 0.148 mmol, 4 mol %),methyl acrylate (0.5 mL, 5.55 mmol) and Et₃N (1.5 mL, 11.1 mmol). Themixture was stirred at 80° C. for 21 h, cooled to rt, and mixed with H₂O(40 mL). The organic compounds were extracted with EtOAc (50 mL), washedwith brine (40 mL), dried (Na₂SO₄), and concentrated. The residue waspurified by flash chromatography to provide 466 mg (47%) of recovered 1cfollowed by 450 mg (49%) of 2c (4,5-Ome).

Preparation PP5

Heck Coupling of 1d and Methyl Acrylate to Form 2d (R=5-NO₂): Theprocedure is same as that of 2c, yielding 82% of 2d after purification.

Preparation PP6 Reductive Amination

Reductive amination of (2a) with benzyl amine to form isoindoline (10a).To a solution of 2a (11.27 g, 59.2 mmol) in ClCH₂CH₂Cl (60 mL) was addedBnNH₂ (6.47 mL, 59.2 mmol), followed by HOAc (5.1 mL, 89 mmol). Themixture was stirred at rt for 1 h. NaCNBH₃ (5.58 g, 88.8 mmol) and MeOH(30 mL) were then added to the above solution. The resulting mixture wasstirred at rt for another 2 h and quenched with sat. NaHCO₃ solution(150 mL). The mixture was extracted with EtOAc (2×100 mL) and thecombined organic layers were washed with brine (150 mL), dried (Na₂SO₄),and concentrated to provide 15.3 g of crude product of 10a which wascarried out for the next hydrogenolysis reaction.

Preparation PP7

One-pot process from 2-carboxaldehyde-methyl-cinnamate to targetcyclized isoindoline product using NaBH₃CN.2-carboxaldehyde-methyl-cinnamate 2a (3.254 g, 17.1 mmol) was dissolvedin a 1:1 MeOH: PhCH₃ mixture (20 mL) at RT. R-(+)-phenethylamine (2.073g, 17.1 mmol) was added and the solution was heated under reflux for 2hours. HPLC in process control indicated that the imine formation wascompleted. Then, AcOH (2.055 g, 34.2 mmol) and NaBH₃CN (2.15 g, 34.2mmol) were successively added at RT, the reaction mixture being cooledwith a water-bath. The reaction mixture was post-agitated overnight.Water (10 mL), MeOH (20 mL) and 37% HCl (2.8 mL) were successively addedand the organic layer was extracted. The aqueous layer was washed withPhCH₃ (10 mL). Then, the aqueous layer was made basic with 5N NaOH (20mL) and MeOH was concentrated to partly remove MeOH. Extraction withEtOAc (2×25 mL) was performed. The combined organic layers were driedover MgSO4, filtered and rinsed with EtOAc (10 mL). The filtrates wereconcentrated under reduced pressure and the residual oil was dried undervacuum overnight at RT to afford the target cyclized isoindoline product10b with 92% yield (4.642 g, 15.7 mmol). HPLC % area indicated that the2 diastereomers were produced in a 55:45 ratio. ¹H NMR confirmed thisresult by integration of the methyl group of the phenethyl substituent.Note: The Heck or Heck-type coupling was performed in toluene with aslight excess of methylacrylate which was removed by distillation beforethe MeOH and the R-(+)-phenethylamine addition.

Preparation PP8

Reductive amination of (2a) with t-butyl carbamate to form (11a): To asolution of aldehyde 2a (238 mg, 1.25 mmol) in CH₃CN (8 mL) was addedt-butyl carbamate (439 mg, 3.75 mmol), followed by triethylsilane (0.6mL, 3.75 mmol) and TEA (0.19 mL, 2.5 mmol). The mixture was stirred atrt overnight, quenched with sat. NaHCO₃ solution (20 mL) and extractedwith EtOAc (2×30 mL). The combined organic layers were washed with brine(30 mL), dried (Na₂SO₄) and concentrated. The residue was purified byflash chromatography (hexane/EtOAc 3:1) to provide 317 mg (87%) of 11a.

Preparation PP9

Reductive amination of 2b with t-butyl carbamate to form 11b: A mixtureof aldehyde 2b (600 mg, 2.72 mmol) Et₃SiH (955 mg, 3 eq, 1.31 mL), TFA(620 mg, 2 eq, 420 uL), t-butyl carbamate (980 mg, 3 eq) in acetonitrile(15 mL) was stirred at room temperature over 2 days. Removed the solventon a Rotary evaporator and purified the crude residue on a flash column(100 g SiO₂, 7:1→6:1 Hex/EtOAc). Collected 307 mg good desired product11b (35%); 195 mg product contaminated with aldehyde SM (22%).

Preparation PP10

Reductive amination of (2c) with t-butyl carbamate to form (11c): To asolution of aldehyde 2c (411 mg, 1.64 mmol) in CH₃CN (10 mL) was addedt-butyl carbamate (580 mg, 4.93 mmol), followed by triethylsilane (0.8mL, 4.93 mmol) and TFA (0.25 mL, 3.28 mmol). The mixture was stirred atrt overnight, quenched with sat. NaHCO₃ solution (30 mL) and extractedwith EtOAc (2×30 mL). The combined organic layers were washed with brine(30 mL), dried (Na₂SO₄) and concentrated. The residue was purified byflash chromatography (hexane/EtOAc 3:1, hexane/EtOAc 1:1) to provide 535mg (93%) of 11c.

Preparation PP11

To a solution of 2d (1.02 g, 4.34 mg) in CH₂Cl₂/CH₃CN (1:1 24 mL) wasadded BocNH₂ (1.5 g, 13.02 mmol), Et₃SiH (2.1 mL, 13.02 mmol), and TFA(0.67 mL, 8.67 mmol). The mixture was stirred at rt for 7 h. Aprecipitate was formed during the reaction. The reaction mixture wasquenched with sat. NaHCO₃ solution (30 mL), and diluted with CH₂Cl₂ (40mL). The organic layer was washed with brine (30 mL), dried (Na₂SO₄),and concentrated. The residue was purified by flash chromatography(hexane/EtOAc 3:1, then CH₂Cl₂/EtOAc 10:1) to provide 2.08 g yellowsolid which still containing BocNH₂. The product is not the desiredBoc-carbamate 14c. LC-MS result showed that the product is the Schiffbase intermediate.

To the above product (420 mg) in CH₂Cl₂ (10 mL) was added Et₃SiH (1 mL)and TFA (0.4 mL). The mixture was stirred at rt for 1 h and small amountof sample was taken for NMR. NMR analysis demonstrated that the startingmaterial was consumed and the product was 14c. TFA (0.7 mL) was thenadded to the above mixture and the resultant solution was stirred at rtfor another 5 h and concentrated. The residue was dissolved in EtOAc (20mL) and washed with H₂O (10 mL). The aqueous layer was basified withsat. NaHCO₃ (30 mL) and the organic compounds were extracted with CH₂Cl₂(2×25 mL). The combined organic layers were washed with brine (20 mL),dried (Na₂SO₄) and concentrated to provide 218 mg of the cyclizedcompound 14c.

Preparation PP12

Condensation of 2a with α-Methylbenzylamine to Form Imine 9.2-carboxaldehyde-methyl-cinnamate 2a (0.897 g, 4.72 mmol) was dissolvedin MeOH (10 mL) at RT. R-(+)-phenethylamine (0.577 g, 4.76 mmol) wasadded and the solution was heated under reflux for 2 hours. HPLC inprocess control indicated that the imine formation was completed. Thesolvent was stripped on a rotary evaporator and the resulting oil wasdried at RT under vacuum overnight. The Schiff base 9 was obtainedalmost quantitatively (1:412 g, 4.81 mmol).

Preparation PP13

Michael Addition

The compound of α-Methyl benzylamine was applied as the auxiliary. Asshown above, the one-pot reaction of aldehyde 2a and α-Methylbenzylamine gave 90% of 10b with a ratio of 1.2:1.

Step-wise Reduction, Amination, and Cyclization

Condensation of aldehyde 2a with α-methylbenzylamine in acetonitrile,methanol, methanol/toluene(1:1) or toluene afforded imine 9 in excellentyield. Reduction of the imine was initially carried out at RT withNaCNBH₃/HOAc. As a result, a poor ee ratio (1.2:1) was obtained,similarly to the previous described one-pot procedure. But when thereaction was carried out with NABH A at RT, the ratio was elevated to2:1. By lowering the reaction temperature to −78° C., the ratio wasincreased to 5 to 6:1.

Preparation PP14

Cyclization of t-Butyl carbamate (11a): The N-Boc isoindoline methylester 12 was originally synthesized from 11a via deprotection of Bocwith TFA, followed by basic workup, and protection with a Boc group.This procedure has been greatly improved by a one-step procedure.

Preparation PP15

In a 3 L 3-neck round bottom flask equipped with a nitrogen inlet,thermocouple and mechanical stirrer, a solution of 160 g (1.15 moles) ofK₂CO₃ in 180 mL of water was stirred at rt. Solid BOC anhydride 120 g(0.55 moles) was added in one portion forming a semi-solution. To thereaction mixture, a solution of the crude amino ester starting material,87 g (0.46 moles) in 120 mL of TBF was added slowly at such a rate tokeep the internal temperature below 35° C. A mild effervescence wasobserved. The reaction mixture was stirred for 18 hours at rt. Analysisof a reaction aliquot via NMR (DMSO₆) indicates the desired product. Thereaction was diluted with brine and the product extracted with EtOAc.The organic layer was dried over Na₂SO₄, filtered, and concentrated toyield a dark oil, 150.1 g, >100% yield. The crude material was taken onto the next step.

Preparation PP16

In a 3-L 3-neck round bottom flask equipped with a mechanical stirrer,thermocouple, and reflux condenser, a solution of 150 g (approx. 0.46moles) of crude N-BOC ester starting material in 750 mL of methanol wasstirred at rt. To the solution, 750 mL of water was added and the cloudymixture was stirred vigorously. Solid LiOH 25 g (1.03 moles) was addedin small portions at such a rate to maintain the internal temperaturebelow 45° C. Upon completion of addition, the reaction was stirredovernight at rt becoming a dark green color. After 18 hours the reactionwas concentrated to yield a thick semisolid. The crude product wasdissolved in EtOAc and washed with 1 N HCl quickly, followed by twobrine washes. The organic layer was dried with Na₂SO₄, filtered andconcentrated to yield 81 g of a dark green solid. The aqueous layerswere combined and back extracted with methylene chloride, dried overNa₂SO₄, filtered, and concentrated to yield 6 g of a dark green solid.Both solids were combined to yield 87 g of desired product confirmed viaNMR (DMSO₆).

Preparation PP17

Synthesis of 14b: Dissolved the N-boc compound 11b (200 mg, 0.62 mmol)in CH₂Cl₂ (1.0 mL). Cooled the clear light yellow solution to 0° C.Added slowly TFA (˜710 mg, 10 eq, ˜500 microliter) via a syringe.Removed the cooling bath and stirred the clear light brown solution atRT overnight. TLC (3:1 Hex/EtOAc, UV) confirmed a complete reaction.Removed the TFA on a rotavapor. Added EtOAc and concentrated again(twice). The crude residue was partitioned between EtOAc (10–15 mL) anda sat. NaHCO₃ (10–15 mL). The aqueous was extracted with EtOAc (2×10mL). The combined organic was dried over MgSO₄, filtered, andconcentrated to yield a light brown wet solid (212 mg, 138%). NMR(CD₃OD) confirmed the desired isoindoline 14b. This crude isoindolinewas used in the next protection step without purification.

Preparation PP18

Synthesis of 12b: To a mixture of the isoindoline 14b (190 mg, 0.859mmol), K₂CO₃ (189 mg, 1.5 eq) in a solvent 1:1 THF/H₂O (1.0 mL) at RTwas added BOC₂O (210 mg, 1.1 eq). The reaction mixture was stirred at RTovernight. TLC (3:1 Hex/EtOAc, UV) confirmed a complete reaction.Diluted the mixture with EtOAc (15 mL), and washed with H₂O (1×20 mL).The aqueous was extracted with EtOAc (1×20 mL). The combined organic waswashed with brine (1×20 mL), dried over MgSO₄, filtered, concentrated toyield a clear brown oil (340 mg, 123%). This crude oil was purified on aprep TALC plate (2×1,000 micron, solvent 2:1.5:0.5 CHCl₃/Hex/EtOAc) toyield 12b a clear yellow oil (190 mg, 69%). ¹H and ¹³C NMR (CDCl₃) wereobtained.

Procedure PP19

Synthesis of 12d (5-NO₂) by Boc-protection. The compound was prepared byfollowing the same procedure as described for 12b.

Preparation PP20

The imine 9 (1.412 g, 4.81 mmol) was dissolved in anhydrous THF (10 mL)at RT and TFA (5 mL) was added. The black solution was then cooled to−78° C. (dry ice bath) and NaBH₄ (0.893 g, 23.6 mmol, 5 eq.) was addedin 2 portions over 5 minutes. Then, the reaction mixture waspost-agitated at −78° C. for 3 hours and allowed to gently warm at RTovernight. Water (20 mL), cyclohexane (10 mL) and EtOH (20 mL) weresuccessively added and the organic layer was extracted and discarded.The aqueous layer was made basic with 5N NaOH (20 mL) and extracted twotimes with a 2:1 EtOAC/PhCH₃ mixture (30 mL). The combined organiclayers were dried over MgSO4, filtered and rinsed with EtOAc (10 mL).The filtrates were concentrated under reduced pressure and the residualoil was dried under vacuum overnight at RT to afford the target cyclizedisoindoline product 10b (1.273 g, 4.31 mmol) with 91.4% yield. HPLC %area indicated that the 2 diastereomers were produced in a 84:16 ratio(de 68%). ¹H NMR confirmed this result by integration of the methylgroup of the phenethyl substituent.

Preparation PP20

N-Boc methyl ester 11a (36.3 g, 0.125 mol) was dissolved in THF (250mL), and the solution was cooled to about 0° C. A solution of potassiumbis(trimethylsilyl) amide (1.24 g, 0.05 mol. eq.) was added slowly via asyringe under nitrogen atmosphere. The temperature was raised about 8degrees during the addition. The cooling bath was removed and thesolution was stirred at r.t. for 30–45 min. The clear brown solution waspoured into a separation funnel containing about 100 mL of a saturatedNH₄Cl. The layers were separated. The aqueous layer was extracted withEtOAc (2×50 mL). The combined organic was washed with brine (1×100 mL),dried over Na₂SO₄, filtered, concentrated on a Rotary evaporator to aclear yellow oil (37.3 g). This crude oil was purified on a flash column(600 g SiO₂), with a gradient solvent 6:1 Hex/EtOAc (2.13 L), 5:1Hex/EtOAc (1.2 L), 4:1 Hex/EtOAc (1.5 L) to yield 12a as a clean yellowoil (34.5 g, 95%).

Preparation PP21

To a solution of 11c (535 mg, 1.52 mmol) in THF (10 mL) was added KHMDS(0.5 M in toluene, 0.1 mL, 0.05 mmol, 2 mol %). The mixture was stirredat r.t. for 20 min, quenched with sat. NH₄Cl solution (20 mL), anddiluted with EtOAc (20 mL). The organic layer was separated, washed withbrine (20 mL), dried (Na₂SO₄) and concentrated. The residue was filteredthrough a plug of silica gel (EtOAc/CH₂Cl₂ 1:10) to give 530 mg (99%) of12c as an off white solid.

Preparation PP22

Deprotections:

Hydrogenolysis of 10a (R=Bn) to Form (14a): To a solution of crude 10a(15.3 g, 54.4 mmol) in MeOH (100 mL) was added Pd(OH)₂/C (Pearlman'scatalyst, 1.02 g, 6 mol %) in a par-shaker bottle. The suspension wasshaken under 30 psi H₂ pressure overnight in the par-shaker, andfiltered through a plug of celite. The filtrate was concentrated toprovide 10.1 g of crude 14a as brown oil. (The procedure is same for themethyl benzylamine isoindoline substrate 10b).

Preparation PP23

In a typical reaction a mixture of the isoindoline ester 12a (92 mg,0.316 mmol) in 1:1 MeOH/H₂O (2 ml) was treated with LiOH (15 mg, 2 eq)at RT overnight. Diluted the mixture with CH₂Cl₂ (5 ml) and water (5ml). Adjusted the pH of the reaction mixture to 1–3 with a 10% NaHSO₄solution. Separated the layers. The aqueous was extracted with CH₂Cl₂(1×10 ml). The combined organic was dried over Na₂SO₄, filtered,concentrated to yield 16a as a pale yellow foam (76 mg, 87%). NMR(CDCl₃) showed a clean desired acid product.

It is noted that he reaction time must be more than 6 hours. The crudefoam can be purified by slurry in warm hexane and then filter to yield atan solid. Hydrolysis using KOH (2–5 eq) in 1:1 MeOH/H₂O overnight wouldgive the same result.

Preparation PP24

Resolution

Purification of Partially Resolved Isoindoline-caboxylic acid methylester: A solution of the crude material (97.62 g) isoindolinecaboxylicacid methyl ester in CH₂Cl₂ (350 mL) was extracted with 1M HCl (400 mL,200 mL). The combined aqueous portions were washed with CH₂Cl₂ (4×250mL) and then made basic with K₂CO₃ solution (85 g in 150 mL of water).The mixture was extracted with CH₂Cl₂ (6×100 mL) and the combinedorganic extracts were dried (Na₂SO₄) and concentrated to give partiallyresolved Isoindolinecaboxylic acid methyl ester as an oil (33.2 g). 60%ee by chiral CE.

Preparation PP25

Resolution of Partially Resolved Isoindoline-caboxylic acid methylester: A solution of partially resolved isoindoline-caboxylic acidmethyl ester (33.24 g, 0.174 mol) in EtOH (130 mL) was treated slowlywith a solution of dibenzoyl-L-tartaric acid (56.06 g, 0.156 mol) inEtOH (200 mL). The solution was seeded with seeded with product andstirred at RT for 4 hours. Pure product was collected by filtration,washed with EtOH (30 mL) and dried to off-white crystals (60.49 g).96.5% ee by chiral CE.

Preparation PP26

Resolution of N-BOC Isoindolinecaboxylic acid: A solution/slurry ofracemic N-BOC Isoindolinecaboxylic acid (114.5 g, 0.413 mol) in EtOAc(1000 mL) was treated slowly with triethylamine (28.8 mL, 0.206 mol),followed by (S)-(−)-α-methylbenzylamine. The solution was seeded withproduct and stirred at RT overnight. The product was collected byfiltration, washed with EtOAc (200 mL) and dried to a white powder(62.98 g). 97.6% ee by chiral CE.

Asymmetric Hydrogenation Routes

Part I: Synthesis of the Z-isomer (Precursor of AsymmetricHydrogenation)

Preparation PP27

Z-isomer 5 was synthesized as outlined in Scheme P1. Compound 5 wasshown to be a single isomer by HPLC and H-1 nmr. The double bondstereochemistry was derived from comparative NOE data using thepurported E-isomer (Scheme P1). The best chiral induction was achievedusing compound 8/Ferrotane/MeOH-THF. With regard to the conversion of 9to 10, which would constitute a formal asymmetric synthesis ofisoindolene 10, this has been achieved using Super hydride-BF₃.OEt_(2.)However, the product was a mixture of 10 and the corresponding de-BOC(deprotected) compound.

Preparation PP28 Compound 2 (Scheme P1)

Phthalic anhydride (751.5 g, 5.014 mole), potassium acetate (498 g,5.014 mole) and acetic anhydride (1 L) were stirred together undernitrogen. The mixture was slowly warmed to 145–150° C. and stirred for10 minutes, then at 140° C. for 20 minutes. The mixture was allowed toslowly cool to 80° C. over 1 hour. Three volumes of water were addedcausing precipitation of a solid. After filtration, the filtered solidwas washed with warm water and pulled as dry as possible for 30 minutes.The solid was then washed with ethanol and acetone respectively. Ifrequired further purification could be achieved by slurring the solid inacetone, at room temperature, for 15 minutes, then filtration. Drying invacuo at 50° C. for 20 hours gave compound 2 as an off-white solid, 470g (48%) with an NMR purity of approx. 90%.

Preparation PP29 Compound 3 (Scheme P1)

Compound 2 (470 g, 2.47 mole) was added to stirred aqueous ammonia (470ml conc. NH₃ in 4.7 L water). The resultant mixture was stirred at roomtemperature for 1 hour then filtered. The filtered solid was washed withwater. The combined aqueous filtrate and washings were carefullyacidified with 6M aq. HCl,(2.35 L). The precipitate was removed byfiltration and dried in vacuo at 50° C. to give compound 3 as a yellowsolid, 259 g (52%).

Preparation PP30 Compound 4 (Scheme P1)

Compound 3 (511 g, 2.7 mole) was slurred in toluene (10 vol). Thionylchloride (385 g, 3.24 mole) was added over 10 minutes to the stirredmixture, which was then heated to reflux for 1.5 hours. H-1 NMR analysisindicated approx. 80% conversion to acid chloride). DMF (3.7 ml) wasadded and the mixture refluxed an additional 3 hours. The resultantmixture was allowed to cool to 35° C. and methanol (1.27 L) added atsuch a rate that the reaction temperature was maintained at 30–35° C.The reaction mixture was kept at this temperature a further 15 minutesthen concentrated in vacuo to give compound 4 as a brown solid, 536 g(quantitative).

Preparation PP31 Compound 5 (Scheme P1)

Compound 4 (750 g, 3.65 mole) was dissolved in acetonitrile (15 L). Thestirred mixture was cooled to 0–5° C. and DMAP (624 g, 5.11 mole) addedin one portion. After 10 minutes BOC anhydride (1115 g, 5.11 mole) wasadded in one portion: there was a slight exotherm accompanied by gasevolution. The mixture was stirred at room temperature for 5 hours, andthen concentrated in vacuo. The residue was dissolved in EtOAc andwashed with 10% aqueous citric acid, satd. aq. Na₂CO₃ and waterrespectively. After drying, concentration of the organics gave a thicksyrup. This material was run through a plug of silica gel (1.5 kg)eluting with 1:1 EtOAc-hexane. Compound 5 was isolated as a dark solid,619 g (55%). Careful chromatography on silica gel eluting with 20%EtOAc-hexane gave 5 as a fluffy white solid.

Scheme P2

Part II: Synthesis of the E-isomer (Precursor of AsymmetricHydrogenation)

Preparation PP32

The E-isomer of Compound 8 (Scheme P2) was prepared as shown in SchemeP2.

Preparation PP33 Compound 7 (Scheme P2)

The compound 7 was prepared according to the procedure of Einhorn et al,Synth. Commun. 2001, 31(5), 741–748.

Preparation PP34 Compound 8 (Scheme P2)

Compound 7 (15.00 g, 60.7 mmole) and methyl(triphenyl phosphoranylidene)acetate (41.40 g, 121.3 mmole) were slurred in toluene (150 ml). Themixture was stirred at reflux and monitored for reaction of 7 by GC.After 1.5 hours the reaction appears complete by GC. After cooling toroom temperature, the mixture was filtered. The solid on the filter waswashed with toluene until colorless. The combined filtrate/washings wereconcentrated in vacuo to leave a tan solid. This material was coated onsilica gel and chromatographed on silica gel (1 kg) eluting with 10%EtOAc-hexane. Compound 8 was isolated as a white or pale yellow powder,5.52 g (30%).

Preparation PP35

Screening of chiral hydrogenation conditions indicated that the bestchiral induction was achieved using compound 8/Ferrotane/MeOH-THF. Withregard to the conversion of 9 to 10, which would constitute a formalasymmetric synthesis of isoindolene 10, this has been achieved usingSuper hydride-BP₃.OEt₂. However, the product was a mixture of 10 and thecorresponding de-BOC (deprotected) compound.

Scheme P4 Coupling of Chiral Isoindoline with d-4-chloro-Phenyl-alanineUsing Tartrate Salt

Preparation PP36 Compound 15 (Scheme P4)

Tartrate salt 14 (58.00 g, 100.27 mmole) was slurred in water (580 ml).Solid NaHCO₃ (25.27 g, 300.8 mmole) was carefully added BOC anhydride(22.98 g, 105.28 mmole) was in one portion and the progress of thereaction monitored by reverse phase HPLC. After 1 hour additional BOCanhydride (2.18 g, 10.00 mmole) was added. The reaction was complete (byHPLC) after 3 hours. The mixture was extracted with EtOAc (2×250 ml.)The combined organic extracts were washed with water (250 ml) and dried(MgSO₄). Filtration and concentration in vacuo gave 15 as a clear lightbrown oil (31.33 g) contaminated with a small amount of t-BuOH and BOCanhydride. This material was used directly in the next reaction.

Preparation PP37 Compound 16 (Scheme P4)

Ester 15 (29.21 g) 100.26 mmole) was dissolved in 3:1 TBF-water (100ml). LiOH (6.00 g, 250.65 mmole) was added in 1 portion to the stirredsolution. After 17 hours, the mixture was stripped to dryness and theresidue was dissolved in water (500 ml.) EtOAc (250 ml) was added andsolid NaHSO₄ added to the stirred mixture until the pH=3. The organiclayer was separated and the aqueous layer extracted with EtOAc (250 ml.)The combined EtOAc layers were dried (MgSO4). Filtration andconcentration in vacuo gave acid 16 as a light tan solid, 27.10 g (97%).

The chemistry used is shown in Scheme P5. Two protocols were used:method A used isolated 16, method B used a solution of 16 derived fromresolved salt 19.

Preparation PP38 Compound 17 (Scheme P5, Method A)

Acid 16 (24.18 g, 87.2 mmole) and D-chloro-phenylalanine hydrochloride(21.81 g, 87.2 mmole) were dissolved in CH₂Cl₂ (100 ml) and DMF (25 ml).The mixture was stirred at ambient temperature. HOBT (13.55 g, 100.3mmole) and Hunig's base (45.6 ml, 33.81 g, 261.6 mmole) were added. HATU(38.13 g, 100.3 mmole) was added in 1 portion (there was a rapidexotherm to 50° C.). The mixture was stirred for 90 minutes then dilutedwith EtOAc (750 ml). The resulting mixture was washed with water, 5%KHSO₄, brine and satd. NaHCO₃ respectively, then dried. Filtration andconcentration in vacuo gave crude 17 as a brown foam. The product waspurified by chromatography on silica gel (1 kg) eluting with 1:1EtOAc-hexane. Ester 17 was isolated as a tan powder, 38.85 g (94%).

Preparation PP39 Compound 17 (Scheme P5, Method B)

Resolved salt 19 (96.27 g, 232.5 mmole) was partitioned between water(500 ml) and CH₂Cl₂ (250 ml) Solid KHSO₄ was added portion wise untilpH=2.5. Separate the organic layer and extract the aqueous layer withCH₂Cl₂ (150 ml). The combined organic layers were dried (MgSO₄) thenfiltered. To this solution was added 4-chloro-D-phenylalanine (58.16 g,232.5 mmole), HOBT (34.57 g, 255.8 mmole), Hunig's base (93.2 ml, 69.13g, 534.9 mmole) and finally HATU (97.26 g, 255.8 mmole). The resultantmixture was stirred at room temperature for 18.5 hours, and then pouredonto a plug of silica gel (1 kg). This was washed with 1:1 EtOAc-hexaneuntil no more product elutes. Ester 17 was isolated as a pink foam,101.79 g (93%): contains about 1% unreacted 16.

Preparation PP40 Compound 18 (Scheme P5)

Ester 17 (38.64 g, 81.7 mmole) was dissolved in 3:1 THF-water (200 ml).LiOH (2.15 g, 89.9 mmole) was added to the mixture, which was stirred atroom temperature for 2 hours. The solvent was then removed in vacuo andthe residual solid taken up in water (600 ml). This was extracted withMTBE (250 ml). The aqueous layer was separated and stirred with EtOAc(250 ml), and solid KHSO₄ was added portion wise until pH=3. The layerswere separated and the aqueous extracted with EtOAc (250 ml). Thecombined organic layers were dried over MgSO4. Filtration andconcentration in vacuo gave acid 18 as a light pink foam, 38.41 g (35.71g corrected for residual solvent, 95%).

Preparation PP41

Step 1: Esterification

In a 22 L 4-neck round bottom flask equipped with a reflux condenser,thermocouple and nitrogen inlet, a slurry of 1000 g (5.4 moles) ofm-tyrosine in 10 L of 2B-3 EtOH was cooled to 5° C. To the slurry, 350ML (12.4 moles) of thionyl chloride were added dropwise via an additionfunnel at such a rate to maintain the reaction temperature below 20° C.Upon completion of addition, the reaction was heated to refluxtemperature and stirred for 18 hrs. The reaction was concentrated toone-third the volume and 8 L of MTBE were charged. The resulting thickslurry was stirred for 14 hrs in a rotary evaporator at r.t. Theresulting solid was isolated on a filter pad and dried at 40° C. for 48hrs yielding 1288 g (95%). NMR (MSOd₆) indicated desired material.

Preparation PP42

Step 2: Pictet-Spengler

In a 22 L 4 neck round bottom flask equipped with a mechanical stirrer,thermocouple, and reflux condenser placed on top of a Soxhlet extractorcharged with 4° A sieves, a semi-solution of m-tyrosine ethyl esterhydrochloride 1288 g (5.26 moles) in 13 L of acetone was heated toreflux temperature. The condensate was filtered through the sieves toremove water. The reaction was stirred vigorously at reflux for 48 hrs.An NMR sample in DMSOd₆ indicated the absence of starting material. Thereaction was cooled to r.t. and concentrated to yield an off-whitesolid, 1411 g (94%).

Preparation PP43

Step 3: Triflation

In a 22 L 4 neck round bottom flask equipped with a reflux condenser,mechanical stirrer, nitrogen inlet, and a thermocouple, 1240 g (4.35moles) of the starting material salt in 12.4 L of methylene chloride wascooled to 4° C. To the mixture, 1452 mL (10.4 moles) of triethylaminewere added and stirred into solution. Triflic anhydride, 1472 mL (5.22moles) was added dropwise to the reaction at such a rate to maintain theinternal temperature below 10° C. The ice bath was removed and thereaction warmed to rt. and stirred for 18 hrs. The reaction wasconcentrated to a oil then dissolved in 4 L of EtOAc and concentratedagain to an oil in an effort to remove excess triflic anhydride Thecrude residue was dissolved in 4 L of EtOAc and washed with water andsaturated sodium bicarbonate solution. The organic layer was isolatedand dried with sodium sulfate, filtered and concentrated to yield 1720 g(>100%) of a crude dark oil which was used without further purification.

Preparation PP44

Step 4: Deoxygenation

A solution of 1720 g (4.35 moles) of crude starting material in 14 L ofacetone was charged to a 10 gallon stainless steel autoclave. To thesolution, a slurry of 5% Pd/C in 1.2 L of toluene was added. Thereaction mixture was evacuated and purged with H₂ gas at 50 psi twotimes. The reaction was stirred overnight at 50° C. with H₂ at 50 psi. Asample aliquot indicated no reaction had occurred. The mixture wasfiltered and concentrated to a thick oil and resubjected to reactionconditions. After 18 hrs, NMR of a sample aliquot indicated absence ofstarting material. The reaction mixture was filtered and the filtrateconcentrated to yield 1581 g of an off-white solid (95%).

Preparation PP45

Step 5: Hydrolysis/Salt Formation

To a 2 L 3 neck round bottom flask equipped with a mechanical stirrer,thermocouple, and nitrogen inlet, a mixture of 700 g (1.83 moles) of thetriflate salt starting material was charged. A solution of 427 g (1.83moles) of the starting material free base in 13.3 L of THF was addedfollowed by 700 mL of water. The semi-solution was stirred vigorously atr.t. To the reaction flask, 43.7 g (1.83 moles) of solid LiOH were addedin small portions at such a rate to maintain the internal temperaturebelow 35° C. The reaction was stirred for 18 hrs at r.t. andconcentrated to yield a thick oil. THF (4 L) was added and thesemi-solution was concentrated. This was repeated with toluene and thesemi-solid was placed under house vacuum on the roto vap with stirringfor 18 hrs to yield 650 g of a crude solid. The solid was reslurried inEtOAc, filtered, isolated and dried to yield 525 g (68%) of the lithiumsalt as an off-white solid.

Preparation PP46

Step 6: Coupling

Solid d-chloro-phenylalanine 446 g (1.78 moles) was added to thesemi-solution followed by 20 g (0.162 moles) of DMAP. The resultingmixture was stirred for 15 minutes then solid EDCl(1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride) 390 g(2.03 moles) was added. The reaction mixture was heated to 80° C. andstirred for 18 hours. Thin layer chromatography (1:1 EtOAc:Hex)indicated very little starting material present. The reaction was cooledto r.t. and concentrated to yield a thick oil. The crude oil wasdissolved in EtOAc and washed with water, and brine. The solution wasdried with sodium sulfate, filtered and concentrated to yield a thickoil, 426 g. The crude oil was chromatographed in several lots using aWaters Prep 500 chromatography apparatus. The eluent consisted of agradient system, 5%–80% EtOAc in heptane at a flow rate of 240 ml/minover 38 minutes. The two diasteromers were separated and isolated toyield 119.04 g for the top spot and 111.3 g for the bottom spot.Conformation of both desired diastereomers was achieved via NMR (DMSO₆).

Preparation PP47 Resolution of Tetrahydroisoquinolinecarboxylic AcidEthyl Ester to Prepare 1-tartaric Acid Salt

Preparation of free-base: A racemic mixture oftetrahydroisoquinolinecarboxylic acid (7.43 g) in EtOAc (60 mL) wastreated with saturated NaHCO₃ solution (60 mL) and saturated Na₂CO₃solution (10 mL). The mixture was agitated and the layers wereseparated. The organic phase was dried (Na₂SO₄) and concentrated to givethe corresponding free-base as an oil (4.85 g)

Resolution: A mixture of the above free base (467 mg, 2.0 mmol), andL-tartaric acid (300 mg, 2.0 mmol) in acetone (4 mL) was stirred at r.t.overnight. The title L-tartaric acid salt was collected by filtration,washed with acetone (about 2 mL) and dried to a white powder (367 mg).100% ee by chiral CE.

Preparation PP48 Resolution of N-BOC TetrahydroisoquinolinecarboxylicAcid

2-{2-[(tert-butyl)oxycarbonyl]-1,2,3,4-tetrahydro-isoquinolyl}aceticacid dehydroabietylamine salt: Racemic2-{2-[(tert-butyl)oxycarbonyl]-1,2,3,4-tetrahydroisoquinolyl}acetic acid(30.15 g, 103.5 mmol) was dissolved in i-PA (300 mL).Dehydroabietylamine (22.11 g, 52.7 mmol of a 68 weight % mixture) wasadded to the solution, which was then agitated on a multi-arm shaker for63 h. The resultant thick paste was filtered and rinsed with i-PA (50mL, 25 mL). Dried in a 50° C. vacuum oven to obtain a white solid (27.73g, 52% ee by chiral CE analysis). The product was reslurried in i-PA(266 mL) and agitated on a multi-arm shaker for 23.5 h. Filtered thethick slurry and rinsed with cold i-PA (50 mL, 30 mL). Dried the cake ina 50° C. vacuum oven and obtained the product as a white solid (23.63 g,40% yield, 94% ee by chiral CE analysis).

Preparation PP49

Enamine 21 (Scheme P6) was prepared as a substrate for asymmetrichydrogenation screening studies. It is formed as an approx. 10:1 mixturewith imine 22. The enamine (21) may be NH-protected i.e., by a Bocprotecting group. The resulting compound 23 may be subjected toasymmetric hydrogenation to afford the acetic acid or methylacetatesubstituted isoquinoline, which may be processed into a compound offormula I as demonstrated previously.

Preparation PP50 Compound 21 (Scheme P6) Prepared as Published W Sobotkaet al, J. Org. Chem., 1965, 30, 3667

Preparation PP51

The chiral synthesis of gem-dimethyl TIC using L-Dopa as the startingmaterial instead of tyrosine was successfully demonstrated up to thePictet-Spengler reaction with L-DOPA and acetone. The product is amixture of starting material 24 and product 25 (major component). Theproduct was isolated by using common isolation procedures. Analternative isolation method is to react the mixture (24 and 25) withBOC anhydride wherein the less hindered N—H in 24 leads to preferentialBOC protection of 24, allowing for ready separation of 25. Chemistry forthe rest of the sequence i.e., deoxygenation reaction, has beendemonstrated herein.

1. A compound of formula I:

or a pharmaceutically acceptable salt or steroisomer thereof, wherein Gis N; A is C₁–C₈ alkyl or C₃–C₇ cycloalkyl; L and L¹ are independently:hydrogen or together oxo; T is:

R is: when y is 1; N(R⁸)₂, NR⁸COR⁸, NR⁸CON(R⁸), NR⁸C(O)OR⁸, NR⁸SO₂R⁸ orR is: when y is 0 or 1; heterocyclyl, provided that when y is 0, aheteroatom is not directly connected to oxygen or adjacent to a carbonthat is connected to oxygen; and wherein the heterocyclyl having atleast one nitrogen in the heterocyclic ring and is optionallysubstituted with one to five substituents independently selected fromR⁸; R¹ is independently: hydrogen, CONH(C₁–C₈ alkyl), C₁–C₈ alkyl,(D)phenyl, (D)C₃–C₇ cycloalkyl or oxo, provided that oxo is not attachedto the ring carbon adjacent to G; R² is independently: hydrogen, haloC₁–C₈ alkyl, C₁–C₈ alkylsulfonyl, (D)C₃–C₇ cycloalkyl or C₁–C₄haloalkyl; R³ is independently: aryl or thienyl; wherein aryl andthienyl are optionally substituted with one to three substituentsselected from the group consisting of: cyano, halo, C₁–C₈ alkyl,(D)C₃–C₇ cycloalkyl, C₁–C₄ alkoxy, C₁–C₄ haloalkyl, benzyloxy, and C₁–C₄haloalkyloxy; R⁴ is independently: hydrogen, C₁–C₈ alkyl, C(O)R⁸,C(O)OR⁸, C₃–C₇ cycloalkyl or (CH₂)_(n)O(C₁–C₈ alkyl), wherein n is 2–8;each R⁸ is independently: hydrogen, phenyl C₁–C₈ alkyl, C₁–C₈alkylsulfonyl, C(O)C₁–C₈ alkyl, C(O)aryl, wherein aryl being phenyl ornaphthyl, SO₂-aryl, wherein aryl being phenyl or naphthyl, (D)C₃–C₇cycloalkyl or (CH₂)_(n)C₁–C₄ haloalkyl, wherein n is 1–8; each R⁹ isindependently: hydrogen, hydroxy, (D)cyano, halo, C₁–C₈ alkyl, C₁–C₈alkoxy, C₃–C₇ cycloalkyl, C₁–C₄ haloalkyl, (D)C(O)R⁸, (D)OR⁸, (D)OCOR⁸,(D)NR⁸SO₂R⁸, each R¹⁰ is independently: hydrogen, (C₁–C₈)alkyl,C(O)C₁–C₈ alkyl, aryl or C₃–C₇ cycloalkyl; each R¹¹ is independently:hydrogen, C₁–C₈ alkyl, (CH₂)_(n)N(R⁸)₂, (CH₂)_(n)NR⁸C(O)C₁–C₄ alkyl, and(CH₂)_(n)NR⁸SO₂C₁–C₄ alkyl, wherein n is 2–8; D is a bond or—(CH₂)_(n)—; n is 0–8; p is 0–4; q is 0–1; y is 0–1.
 2. The compound ofclaim 1, wherein the R heterocyclyl is a 4-, 5- or 6-membered ringhaving one nitrogen atom.
 3. The compound of claim 2, wherein the Rheterocyclyl is a 6-membered ring having one nitrogen and one oxygenatom.
 4. The compound of claim 3, wherein R³ is phenyl optionallypara-substituted with fluoro, chloro, bromo, iodo, benzyloxy, methoxy ormethyl.
 5. The compound of claim 4, wherein R³ is phenylpara-substituted with chloro, fluoro or methoxy.
 6. The compound ofclaim 5, wherein R⁴ is hydrogen.
 7. The compound of claim 6, wherein—(CH₂)_(n)—T is:

where * denotes a chiral carbon atom having a R or S configuration. 8.The compound of claim 7, wherein L and L¹ are together oxo and thechiral carbon has R configuration.
 9. A compound of formula II,

or a pharmaceutically acceptable salt or steroisomer thereof, wherein Ais C₁–C₈ alkyl or C₃–C₇ cycloalkyl; r is 0 or 1; y is 0 or 1; D is abond or —(CH₂)_(n)—; n is 0–8; R is: when y is 1; N(R⁸)₂, NR⁸COR⁸,NR⁸CON(R⁸)₂, NR⁸C(O)OR⁸, NR⁸SO₂R⁸ or R is: when y is 0 or 1;heterocyclyl, provided that when y is 0, a heteroatom is not directlyconnected to oxygen or adjacent to a carbon that is connected to oxygen;and wherein the heterocyclyl has at least one nitrogen in theheterocyclic ring and is optionally substituted with one to fivesubstituents independently selected from R⁸; each R⁸ is independently:hydrogen, phenyl C₁–C₈ alkyl, C₁–C₈ alkylsulfonyl, C(O)C₁–C₈ alkyl,C(O)aryl, wherein aryl being phenyl or naphthyl, SO₂-aryl, wherein arylbeing phenyl or naphthyl, (D)C₃–C₇ cycloalkyl or (CH₂)_(n)C₁C₄haloalkyl, wherein n is 1–8; each R¹⁰ is independently: hydrogen,(C₁–C₈)alkyl, C(O)C₁–C₈ alkyl, aryl or C₃–C₇ cycloalkyl.
 10. Thecompound of claim 9, wherein O—(A)_(y)—R is attached to the orthoposition of the phenyl ring.
 11. The compound of claim 10, wherein the Rheterocyclyl is a 4-, 5- or 6-membered ring having one nitrogen atom.12. The compound of claim 11, wherein the nitrogen is substituted withone substituent selected from R⁸ when y is
 0. 13. The compound of claim11, wherein the R heterocyclyl is a 6-membered ring having one nitrogenand one oxygen atom.
 14. A compound of formula IV,

or a pharmaceutically acceptable salt or steroisomer thereof, wherein Ais C₁–C₈ alkyl or C₃–C₇ cycloalkyl; r is 0 or 1; y is 0 or 1; D is abond or —(CH₂)_(n)—; n is 0–8; R is: when y is 1; N(R⁸)₂, NR⁸COR⁸,NR⁸CON(R⁸)₂, NR⁸C(O)OR⁸, NR⁸SO₂R⁸ or R is: when y is 0 or 1;heterocyclyl, provided that when y is 0, a heteroatom is not directlyconnected to oxygen or adjacent to a carbon that is connected to oxygen;and wherein the heterocyclyl has at least one nitrogen in theheterocyclic ring and is optionally substituted with one to fivesubstituents independently selected from R⁸; each R⁸ is independently:hydrogen, phenyl C₁–C₈ alkyl, C₁–C₈ alkylsulfonyl, C(O)C₁–C₈ alkyl,C(O)aryl, wherein aryl being phenyl or naphthyl, SO₂-aryl, wherein arylbeing phenyl or naphthyl, (D)C₃–C₇ cycloalkyl or (CH₂)_(n)C₁–C₄haloalkyl, wherein n is 1–8; each R¹⁰ is independently: hydrogen,(C₁–C₈)alkyl, C(O)C₁–C₈ alkyl, aryl or C₃–C₇ cycloalkyl.
 15. Thecompound of claim 14, wherein O—(A)_(y)—R is attached to the orthoposition of the phenyl ring.
 16. The compound of claim 15, wherein the Rheterocyclyl is a 4-, 5- or 6-membered ring having one nitrogen atom.17. The compound of claim 16, wherein the nitrogen is substituted withone substituent selected from R⁸ when y is
 0. 18. The compound of claim16, wherein the heterocyclyl is a 6-membered ring having one nitrogenand one oxygen atom.
 19. A compound of formula V,

or a pharmaceutically acceptable salt or steroisomer thereof, wherein Ais C₁–C₈ alkyl or C₃–C₇ cycloalkyl; y is 0 or 1; D is a bond or—(CH₂)_(n)—; n is 0–8; R is: when y is 1; N(R⁸)₂, NR⁸COR⁸, NR⁸CON(R⁸)₂,NR⁸C(O)OR⁸, NR⁸SO₂R⁸ or R is: when y is 1 or 1; heterocyclyl, providedthat when y is 0, a heteroatom is not directly connected to oxygen oradjacent to a carbon that is connected to oxygen; and wherein theheterocyclyl has at least one nitrogen in the heterocyclic ring and isoptionally substituted with one to five substituents independentlyselected from R⁸; each R⁸ is independently: hydrogen, phenyl C₁–C₈alkyl, C₁–C₈ alkylsulfonyl, C(O)C₁–C₈ alkyl, C(O)aryl, wherein arylbeing phenyl or naphthyl, SO₂-aryl, wherein aryl being phenyl ornaphthyl, (D)C₃–C₇ cycloalkyl or (CH₂)_(n)C₁–C₄ haloalkyl, wherein n is1–8.
 20. The compound of claim 19, wherein O—(A)_(y)—R is attached tothe ortho position of the phenyl ring.
 21. The compound of claim 20,wherein the R heterocyclyl is a 4-, 5- or 6-membered ring having onenitrogen atom.
 22. The compound of claim 21, wherein the nitrogen issubstituted with one substituent selected from R⁸ when y is
 0. 23. Thecompound of claim 21, wherein the heterocyclyl is a 6-membered ringhaving one nitrogen and one oxygen atom.
 24. A compound of formula VI,

or a pharmaceutically acceptable salt or steroisomer thereof, wherein Ais C₁–C₈ alkyl or C₃–C₇ cycloalkyl; y is 0 or 1; D is a bond or—(CH₂)_(n)—; n is 0–8; R is: when y is 1; N(R⁸)₂, NR⁸COR⁸, NR⁸CON(R⁸)₂,NR⁸C(O)OR⁸, NR⁸SO₂R⁸ or R is: when y is 0 or 1; heterocyclyl, providedthat when y is 0, a heteroatom is not directly connected to oxygen oradjacent to a carbon that is connected to oxygen; and wherein theheterocyclyl has at least one nitrogen in the heterocyclic ring and isoptionally substituted with one to five substituents independentlyselected from R⁸; each R⁸ is independently: hydrogen, phenyl C₁–C₈alkyl, C₁–C₈ alkylsulfonyl, C(O)C₁–C₈ alkyl, C(O)aryl, wherein arylbeing phenyl or naphthyl, SO₂-aryl, wherein aryl being phenyl ornaphthyl, (D)C₃–C₇ cycloalkyl or (CH₂)_(n)C₁–C₄ haloalkyl, wherein n is1–8.
 25. The compound of claim 24, wherein O—(A)_(y)—R is attached tothe ortho position of the phenyl ring.
 26. The compound of claim 25,wherein the R heterocyclyl is a 4-, 5- or 6-membered ring having onenitrogen atom.
 27. The compound of claim 26, wherein the nitrogen issubstituted with one substituent selected from R⁸ when y is
 0. 28. Thecompound of claim 26, wherein the heterocyclyl is a 6-membered ringhaving one nitrogen and one oxygen atom.
 29. A compound selected fromthe group consisting of: Name of Compound Compound(N-(1-(4-R-chlorobenzyl)-2-{4-[2-(1-methyl-S-piperidin-3-yloxy)-phenyl]-piperazin-1-yl}-2-oxo-ethyl)-2-(2,3-dihydro-1H-isoindol-1-yl)-aceamide,trihydrochloride

(N-(1-(4-R-chlorobenzyl)-2-oxo-2-{4-[2-(R-piperidin-3-yloxy)-phenyl]-piperazin-1-yl}ethyl)-2-(2,3-dihydro-1H-isoindol-1-yl)-aceamide,trihydrochloride

2-(2,3-dihydro-1H-isoindol-1-yl)-N-(1-(4-methoxy-benzyl)-2-{4-[2-(1-methyl-piperidin-3-yloxy)-phenyl]-piperazin-1-yl)-2-oxo-ethyl)-acetaimde,trihydrochloride


30. A pharmaceutical composition which comprises a pharmaceuticalcarrier and at least one compound of formula I or its pharmaceuticallyacceptable salts or stereoisomer thereof as recited in claim
 1. 31. Aprocess of making a pharmaceutical composition comprising a compound offormula I or a pharmaceutically acceptable salt or stereoisomers thereofas recited in claim 1 and a pharmaceutically acceptable carrier.
 32. Amethod of treating obesity in a mammal comprising the administration ofa therapeutically effective amount of the compound of formula I asrecited in claim
 1. 33. A process for preparing a compound of formula I:

or a pharmaceutically acceptable salt or steroisomer thereof, wherein Gis N; A is C₁–C₈ alkyl or C₃–C₇ cycloalkyl; —CLL¹—(CH₂)_(n)—T is:

wherein R¹ is hydrogen, C₁–C₈ alkyl, Boc, CBZ, FMOC, phenyl or (C₁–C₈alkyl)phenyl; Q represents a moiety:

R is: when y is 1; N(R⁸)₂, NR⁸COR⁸, NR⁸CON(R⁸)₂, NR⁸C(O)OR⁸, NR⁸SO₂R⁸ orR is: when y is 0 or 1; heterocyclyl, provided that when y is 0, aheteroatom is not directly connected to oxygen or adjacent to a carbonthat is connected to oxygen; and wherein the heterocyclyl has at leastone nitrogen in the heterocyclic ring and is optionally substituted withone to five substituents independently selected from R⁸; R¹ isindependently: hydrogen, CONH(C₁–C₈ alkyl), C₁–C₈ alkyl, (D)phenyl,(D)C₃–C₇ cycloalkyl or oxo, provided that oxo is not attached to thering carbon adjacent to G; R² is independently: hydrogen, halo C₁–C₈alkyl, C₁–C₈ alkylsulfonyl, (D)C₃–C₇ cycloalkyl or C₁–C₄ haloalkyl; R³is independently: aryl or thienyl; wherein aryl and thienyl areoptionally substituted with one to three substituents selected from thegroup consisting of: cyano, halo, C₁–C₈ alkyl, (D)C₃–C₇ cycloalkyl,C₁–C₄ alkoxy, C₁–C₄ haloalkyl, benzyloxy, and C₁–C₄ haloalkyloxy; R⁴ isindependently: hydrogen, C₁–C₈ alkyl, C(O)R⁸, C(O)OR⁸, C₃–C₇ cycloalkylor (CH₂)_(n)O(C₁–C₈ alkyl), wherein n is 2–8; each R⁸ is independently:hydrogen, phenyl C₁–C₈ alkyl, C₁–C₈ alkylsulfonyl, C(O)C₁–C₈ alkyl,C(O)aryl, wherein aryl being phenyl or naphthyl, SO₂-aryl, wherein arylbeing phenyl or naphthyl, (D)C₃–C₇ cycloalkyl or (CH₂)_(n)C₁–C₄haloalkyl, wherein n is 1–8; each R⁹ is independently: hydrogen,hydroxy, (D)cyano, halo, C₁–C₈ alkyl, C₁–C₈ alkoxy, C₃–C₇ cycloalkyl,C₁–C₄ haloalkyl, (D)C(O)R⁸, (D)OR⁸, (D)OCOR⁸, each R¹⁰ is independently:hydrogen, (C₁–C₈)alkyl, C(O)C₁–C₈ alkyl, aryl or C₃–C₇ cycloalkyl; D isa bond or —(CH₂)_(n)—; n is 0–8; p is 0–4; q is 0–1; and y is 0–1;comprising the steps of: a) reacting a compound having a structuralformula 1,

 with CH₂CH═C(O)OR^(a) wherein R^(a) is hydrogen or C₁–C₈ alkyl and X ishalo, in the presence of a catalyst and a base in a suitable organicsolvent to give the compound of formula 2,

b) reductively aminating the compound of formula 2 in the presence ofamine in an acidic condition to give a compound of formula 3,

c) cyclizing the compound of formula 3 by Michael addition to give acompound of formula 4 or stereoisomers thereof,

d) coupling the compound of formula 4 or stereoisomers thereof, whereinR^(a) of compound 4 is H, with a compound of formula 5,

 wherein R^(a) of compound 5 is C₁–C₈ alkyl, to give a compound offormula 6;

e) coupling the compound of formula 6, wherein R^(a) is H, with acompound having a structural,

 to afford the compound of formula
 1. 34. The process of claim 33,wherein

in Step (a) is 2-bromobenzaldehydes.
 35. The process of claim 33,wherein CH₂CH═C(O)OR^(a) in Step (a) is methylacrylate.
 36. The processof claim 35, wherein the catalyst in Step (a) is selected from the groupconsisting of: Pd(Ph₃P)₂Cl₂, Pd(Ph₃P)₄Cl₂, Pd(Ph₃P)₄, Pd(Ph₃ P)₂Cl₂/CuI,Pd(OAc)₂/Ph₃P—Bu₄NBr, Pd(Ph₃P)₄Cl₂/H₂ and Pd(OAc)₂/P(O-tol)₃; andwherein the base in Step (a) is NR₃ wherein R is hydrogen or C₁–C₈alkyl.
 37. The process of claim 36, wherein the amine in Step (b) isselected from the group consisting of: benzylamine,alpha-methylbenzylamine and BocNH₂.
 38. The process of claim 36, whereinthe Step (b) further comprises reducing an incipient imine compound inthe presence of reducing agent, the reducing agent being selected fromthe group consisting of: NaCNBH₃, Na(OAc)₃BH, NaBH₄/H+, and acombination of Et₃SiH and TFA in CH₃CN or CH₂Cl₂.
 39. The process ofclaim 38, wherein the stereoisomer of compound of formula 4 in Step (c)is a compound of formula 4a


40. The process of claim 38, wherein the compound of formula 4a isprepared by asymmetric hydrogenation of a compound having structuralformula,


41. The process of claim 40, wherein the Michael addition in Step (c) iscarried out in a basic workup condition.
 42. The process of claim 33,wherein the Step (e) further comprises deprotecting or protecting of thecompound of formula (6) at NR₁.